Inhibitors of serine proteases, particularly HCV NS3-NS4A protease

ABSTRACT

The present invention relates to compounds of formula I or formula Ia or pharmaceutically acceptable salts thereof, that inhibit serine protease activity, particularly the activity of hepatitis C virus NS3-NS4A protease. As such, they act by interfering with the life cycle of the hepatitis C virus and are useful as antiviral agents. The invention further relates to pharmaceutically acceptable compositions comprising said compounds either for ex vivo use or for administration to a patient suffering from HCV infection and processes for preparing the compounds. The invention also relates to methods of treating an HCV infection in a patient by administering a pharmaceutical composition comprising a compound of this invention.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of co-pending U.S. patent applicationSer. No. 10/893,748, filed Jul. 16, 2004, entitled “Inhibitors of SerineProteases, Particularly HCV NS3-NS4A Protease”, which claims the benefitof U.S. Provisional Patent Application 60/488,535, filed Jul. 18, 2003,entitled “Inhibitors of Serine Proteases, Particularly HCV NS3-NS4AProtease”, both of which are incorporated in their entirety herein byreference.

TECHNICAL FIELD OF THE INVENTION

The present invention relates to compounds that inhibit serine proteaseactivity, particularly the activity of hepatitis C virus NS3-NS4Aprotease. As such, they act by interfering with the life cycle of thehepatitis C virus and are also useful as antiviral agents. The inventionfurther relates to pharmaceutical compositions comprising thesecompounds either for ex vivo use or for administration to a patientsuffering from HCV infection. The invention also relates to processesfor preparing the compounds and methods of treating an HCV infection ina patient by administering a pharmaceutical composition comprising acompound of this invention.

BACKGROUND OF THE INVENTION

Infection by hepatitis C virus (“HCV”) is a compelling human medicalproblem. HCV is recognized as the causative agent for most cases ofnon-A, non-B hepatitis, with an estimated human sero-prevalence of 3%globally [A. Alberti et al., “Natural History of Hepatitis C,” J.Hepatology, 31., (Suppl. 1), pp. 17-24 (1999)]. Nearly four millionindividuals may be infected in the United States alone [M. J. Alter etal., “The Epidemiology of Viral Hepatitis in the United States,Gastroenterol. Clin. North Am., 23, pp. 437-455 (1994); M. J. Alter“Hepatitis C Virus Infection in the United States,” J. Hepatology, 31.,(Suppl. 1), pp. 88-91 (1999)].

Upon first exposure to HCV only about 20% of infected individualsdevelop acute clinical hepatitis while others appear to resolve theinfection spontaneously. In almost 70% of instances, however, the virusestablishes a chronic infection that persists for decades [S. Iwarson,“The Natural Course of Chronic Hepatitis,” FEMS Microbiology Reviews,14, pp. 201-204 (1994); D. Lavanchy, “Global Surveillance and Control ofHepatitis C,” J. Viral Hepatitis, 6, pp. 35-47 (1999)]. This usuallyresults in recurrent and progressively worsening liver inflammation,which often leads to more severe disease states such as cirrhosis andhepatocellular carcinoma [M. C. Kew, “Hepatitis C and HepatocellularCarcinoma”, FEMS Microbiology Reviews, 14, pp. 211-220 (1994); I. Saitoet. al., “Hepatitis C Virus Infection is Associated with the Developmentof Hepatocellular Carcinoma,” Proc. Natl. Acad. Sci. USA, 87, pp.6547-6549 (1990)]. Unfortunately, there are no broadly effectivetreatments for the debilitating progression of chronic HCV.

HCV is a RNA virus of the Flaviviridae family. Acute infection with HCVcauses a generally mild, often asymptomatic, acute hepatitis. However,at least 85% of patients infected with HCV do not fully clear the virusand develop chronic infection of the liver. Once chronic hepatitis C isestablished, spontaneous clearance of the virus is rare and the majorityof patients with chronic hepatitis C develop slowly progressive liverdisease. Twenty years after infection, most patients have evidence ofongoing chronic hepatitis and at least 20% have cirrhosis. Long-termsequelae of chronic hepatitis C include cirrhosis, hepatic failure, andhepatocellular carcinoma. It is estimated that HCV infects 170 millionpersons worldwide. Over the next ten years, as a larger proportion ofpatients who are currently infected enter the third decade of theirinfection, the number of deaths attributed to hepatitis C is expected tosignificantly increase.

Typical symptoms of HCV infection include elevated ALT, positive testfor anti-HCV antibodies, presence of HCV as demonstrated by a positivetest for HCV-RNA, clinical stigmata of chronic liver disease, orhepatocellular damage.

The HCV genome encodes a polyprotein of 3010-3033 amino acids [Q. L.Choo, et. al., “Genetic Organization and Diversity of the Hepatitis CVirus.” Proc. Natl. Acad. Sci. USA, 88, pp. 2451-2455 (1991); N. Kato etal., “Molecular Cloning of the Human Hepatitis C Virus Genome FromJapanese Patients with Non-A, Non-B Hepatitis,” Proc. Natl. Acad. Sci.USA, 87, pp. 9524-9528 (1990); A. Takamizawa et. al., “Structure andOrganization of the Hepatitis C Virus Genome Isolated From HumanCarriers,” J. Virol., 65, pp. 1105-1113 (1991)]. The HCV nonstructural(NS) proteins are presumed to provide the essential catalytic machineryfor viral replication. The NS proteins are derived by proteolyticcleavage of the polyprotein [R. Bartenschlager et. al., “NonstructuralProtein 3 of the Hepatitis C Virus Encodes a Serine-Type ProteinaseRequired for Cleavage at the NS3/4 and NS4/5 Junctions,” J. Virol., 67,pp. 3835-3844 (1993); A. Grakoui et. al., “Characterization of theHepatitis C Virus-Encoded Serine Proteinase: Determination ofProteinase-Dependent Polyprotein Cleavage Sites,” J. Virol., 67, pp.2832-2843 (1993); A. Grakoui et. al., “Expression and Identification ofHepatitis C Virus Polyprotein Cleavage Products,” J. Virol., 67, pp.1385-1395 (1993); L. Tomei et. al., “NS3 is a serine protease requiredfor processing of hepatitis C virus polyprotein”, J. Virol., 67, pp.4017-4026 (1993)].

The HCV NS protein 3 (NS3) contains a serine protease activity thathelps process the majority of the viral enzymes, and is thus consideredessential for viral replication and infectivity. It is known thatmutations in the yellow fever virus NS3 protease decrease viralinfectivity [Chambers, T. J. et. al., “Evidence that the N-terminalDomain of Nonstructural Protein NS3 From Yellow Fever Virus is a SerineProtease Responsible for Site-Specific Cleavages in the ViralPolyprotein”, Proc. Natl. Acad. Sci. USA, 87, pp. 8898-8902 (1990)]. Thefirst 181 amino acids of NS3 (residues 1027-1207 of the viralpolyprotein) have been shown to contain the serine protease domain ofNS3 that processes all four downstream sites of the HCV polyprotein [C.Lin et al., “Hepatitis C Virus NS3 Serine Proteinase: Trans-CleavageRequirements and Processing Kinetics”, J. Virol., 68, pp. 8147-8157(1994)].

The HCV NS3 serine protease and its associated cofactor, NS4A, helpsprocess all of the viral enzymes, and is thus considered essential forviral replication. This processing appears to be analogous to thatcarried out by the human immunodeficiency virus aspartyl protease, whichis also involved in viral enzyme processing. HIV protease inhibitors,which inhibit viral protein processing, are potent antiviral agents inman, indicating that interrupting this stage of the viral life cycleresults in therapeutically active agents. Consequently HCV NS3 serineprotease is also an attractive target for drug discovery.

Furthermore, the current understanding of HCV has not led to any othersatisfactory anti-HCV agents or treatments. Until recently, the onlyestablished therapy for HCV disease was interferon treatment (see, e.g.,PCT publication No. WO 02/18369, the disclosure of which is hereinincorporated by reference). However, interferons have significant sideeffects [M. A. Wlaker et al., “Hepatitis C Virus: An Overview of CurrentApproaches and Progress,” DDT, 4, pp. 518-29 (1999); D. Moradpour etal., “Current and Evolving Therapies for Hepatitis C,” Eur. J.Gastroenterol. Hepatol., 11, pp. 1199-1202 (1999); H. L. A. Janssen etal. “Suicide Associated with Alfa-Interferon Therapy for Chronic ViralHepatitis,” J. Hepatol., 21, pp. 241-243 (1994); P. F. Renault et al.,“Side Effects of Alpha Interferon,” Seminars in Liver Disease, 9, pp.273-277. (1989)] and induce long term remission in only a fraction(˜25%) of cases [O. Weiland, “Interferon Therapy in Chronic Hepatitis CVirus Infection”, FEMS Microbiol. Rev., 14, pp. 279-288 (1994)].Ribavirin, a broad spectrum antiviral agent, has reported acitivty inchronic hepatitis C. Recent introductions of the pegylated forms ofinterferon (PEG-Intron® and Pegasys®) and the combination therapy ofribavirin and pegylated interferon (Rebetrol®) have resulted in onlymodest improvements in remission rates and only partial reductions inside effects (see, e.g., U.S. Pat. No. 6,299,872, U.S. Pat. No.6,387,365, U.S. Pat. No. 6,172,046, U.S. Pat. No. 6,472,373, thedisclosures of which are incorporated herein by reference). Moreover,the prospects for effective anti-HCV vaccines remain uncertain.

Thus, there is a need for more effective anti-HCV therapies particularlycompounds that may be used as protease inhibitors. Such inhibitors wouldhave therapeutic potential as protease inhibitors, particularly asserine protease inhibitors, and more particularly as HCV NS3 proteaseinhibitors. Specifically, such compounds may be useful as antiviralagents, particularly as anti-HCV agents.

The present invention provides compounds that are potent binders andinhibitors of the HCV NS3/NS4A serine protease and are, therefore,useful as anti-HCV agents.

SUMMARY OF THE INVENTION

The present invention provides a compound of formula I:

or a pharmaceutically acceptable salt thereof,wherein:

-   X and X′ are both fluorine; or-   X and X′ are independently C(H), N, NH, O, or S; and X and X′ are    taken together with the carbon atom to which they are bound to form    a 5- to 7-membered saturated or partially unsaturated ring having up    to 4 heteroatoms independently selected from N, NH, O, S, SO, and    SO₂; wherein any atom is optionally singly or multiply substituted    with up to 3 substituents selected independently from J; and wherein    said ring is optionally fused to a second ring selected from    (C6-C10)aryl, (C5-C10)heteroaryl, (C3-C10)cycloalkyl, and a    (C3-C10)heterocyclyl, wherein said second ring has up to 3    substituents selected independently from J;    -   J is halogen, —OR′, —NO₂, —CN, —CF₃, —OCF₃, —R′, oxo, thioxo,        ═N(R′), ═N(OR′), 1,2-methylenedioxy, 1,2-ethylenedioxy, —N(R′)₂,        —SR′, —SOR′, —SO₂R′, —SO₂N(R′)₂, —SO₃R′, —C(O)R′, —C(O)C(O)R′,        —C(O)C(O)OR′, —C(O)C(O)NR′, —C(O)CH₂C(O)R′, —C(S)R′, —C(S)OR′,        —C(O)OR′, —OC(O)R′, —C(O)N(R′)₂, —OC(O)N(R′)₂, —C(S)N(R′)₂,        —(CH₂)₀₋₂NHC(O)R′, —N(R′)N(R′)COR′, —N(R′)N(R′)C(O)OR′,        —N(R′)N(R′)CON(R′)₂, —N(R′)SO₂R′, —N(R′)SO₂N(R′)₂,        —N(R′)C(O)OR′, —N(R′)C(O)R′, —N(R′)C(S)R′, —N(R′)C(O)N(R′)₂,        —N(R′)C(S)N(R′)₂, —N(COR′)COR′, —N(OR′)R′, —C(═NH)N(R′)₂,        —C(O)N(OR′)R′, —C(═NOR′)R′, —OP(O)(OR′)₂, —P(O)(R′)₂,        —P(O)(OR′)₂, or —P(O)(H)(OR′); wherein;        -   R′ is independently selected from:        -   hydrogen-,        -   (C1-C12)-aliphatic-,        -   (C3-C10)-cycloalkyl- or -cycloalkenyl-,        -   [(C3-C10)-cycloalkyl or -cycloalkenyl]-(C1-C12)-aliphatic-,        -   (C6-C10)-aryl-,        -   (C6-C10)-aryl-(C1-C12)aliphatic-,        -   (C3-C10)-heterocyclyl-,        -   (C3-C10)-heterocyclyl-(C1-C12)aliphatic-,        -   (C5-C10)-heteroaryl-, and        -   (C5-C10)-heteroaryl-(C1-C12)-aliphatic-;        -   wherein up to 5 atoms in R′ are optionally and independently            substituted with J;        -   wherein two R′ groups bound to the same atom form a 3- to            10-membered aromatic or non-aromatic ring having up to 3            heteroatoms independently selected from N, NH, O, S, SO, and            SO₂, wherein said ring is optionally fused to a            (C6-C10)aryl, (C5-C10)heteroaryl, (C3-C10)cycloalkyl, or a            (C3-C10)heterocyclyl, wherein any ring has up to 3            substituents selected independently from J;-   Y and Y′ are independently:    -   hydrogen-,    -   (C1-C12)-aliphatic-,    -   (C3-C10)-cycloalkyl- or -cycloalkenyl-,    -   (C3-C10)-cycloalkyl-(C1-C12)-aliphatic-,    -   (C6-C10)-aryl-,    -   (C3-C10)-heterocyclyl-; or    -   (C5-C10)-heteroaryl-;        -   wherein up to three aliphatic carbon atoms in Y and Y′ may            be replaced by O, N, NH, S, SO, or SO₂;        -   wherein each of Y and Y′ is independently and optionally            substituted with up to 3 substituents independently selected            from J;-   R₁ and R₃ are independently:    -   (C1-C12)-aliphatic-,    -   (C3-C10)-cycloalkyl- or -cycloalkenyl-,    -   [(C3-C10)-cycloalkyl- or -cycloalkenyl]-(C1-C12)-aliphatic-,    -   (C6-C10)-aryl-(C1-C12)aliphatic-, or    -   (C5-C10)-heteroaryl-(C1-C12)-aliphatic-;        -   wherein up to 3 aliphatic carbon atoms in R₁ and R₃ may be            replaced by a heteroatom selected from O, N, NH, S, SO, or            SO₂ in a chemically stable arrangement;        -   wherein each of R₁ and R₃ is independently and optionally            substituted with up to 3 substituents independently selected            from J;-   R₂, R₄, and R₇ are independently:    -   hydrogen-,    -   (C1-C12)-aliphatic-,    -   (C3-C10)-cycloalkyl-(C1-C12)-aliphatic-, or    -   (C6-C10)-aryl-(C1-C12)-aliphatic-;        -   wherein up to two aliphatic carbon atoms in R₂, R₄, and R₇            may be replaced by a heteroatom selected from O, N, NH, S,            SO, and SO₂ in a chemically stable arrangement;        -   wherein each of R₂, R₄, and R₇ is independently and            optionally substituted with up to 3 substituents            independently selected from J;-   R₅ and R_(5′) are independently hydrogen or (C1-C12)-aliphatic,    wherein any hydrogen is optionally replaced with halogen; wherein    any terminal carbon atom of R₅ is optionally substituted with    sulfhydryl or hydroxy; or R₅ is Ph or —CH₂Ph and R_(5′), is H,    wherein said Ph or —CH₂Ph group is optionally substituted with up to    3 substituents independently selected from J; or-   R₅ and R_(5′) together with the atom to which they are bound is a 3-    to 6-membered saturated or partially unsaturated ring having up to 2    heteroatoms selected from N, NH, O, SO, and SO₂; wherein the ring    has up to 2 substituents selected independently from J;-   W is:

-   -   wherein each R₆ is independently:        -   hydrogen-,        -   (C1-C12)-aliphatic-,        -   (C6-C10)-aryl-,        -   (C6-C10)-aryl-(C1-C12)aliphatic-,        -   (C3-C10)-cycloalkyl- or cycloalkenyl-,        -   [(C3-C10)-cycloalkyl- or cycloalkenyl]-(C1-C12)-aliphatic-,        -   (C3-C10)-heterocyclyl-,        -   (C3-C10)-heterocyclyl-(C1-C12)-aliphatic-,        -   (C5-C10)-heteroaryl-, or        -   (C5-C10)-heteroaryl-(C1-C12)-aliphatic-, or        -   two R₆ groups, which are bound to the same nitrogen atom,            form together with that nitrogen atom, a            (C3-C10)-heterocyclic ring;        -   wherein R₆ is optionally substituted with up to 3 J            substituents;    -   wherein each R₈ is independently —OR′; or the R₈ groups together        with the boron atom, is a (C3-C10)-membered heterocyclic ring        having in addition to the boron up to 3 additional heteroatoms        selected from N, NH, O, SO, and SO₂;

-   V is O or a valence bond; and

-   T is:    -   (C1-C12)-aliphatic-;    -   (C6-C10)-aryl-,    -   (C6-C10)-aryl-(C1-C12)aliphatic-,    -   (C3-C10)-cycloalkyl or -cycloalkenyl-,    -   [(C3-C10)-cycloalkyl or -cycloalkenyl]-(C1-C12)-aliphatic-,    -   (C3-C10)-heterocyclyl-,    -   (C3-C10)-heterocyclyl-(C1-C12)-aliphatic-,    -   (C5-C10)-heteroaryl-, or    -   (C5-C10)-heteroaryl-(C1-C12)-aliphatic-;        -   wherein up to 3 aliphatic carbon atoms in T may be replaced            by a heteroatom selected from O, N, NH, S, SO, or SO₂ in a            chemically stable arrangement;        -   wherein each T is optionally substituted with up to 3 J            substituents;            provided that the following compounds are excluded:

-   a)    N-acetyl-L-leucyl-(2S)-2-cyclohexylglycyl-(3S)-6,10-dithia-2-azaspiro[4.5]decane-3-carbonyl-3-amino-2-oxohexanoylglycyl-2-phenyl-1,1-dimethylethyl    ester glycine;

-   b)    N-acetyl-L-leucyl-(2S)-2-cyclohexylglycyl-(3S)-6,10-dithia-2-azaspiro[4.5]decane-3-carbonyl-3-amino-2-oxohexanoylglycyl-2-phenyl-glycine;

-   c)    N-acetyl-L-leucyl-(2S)-2-cyclohexylglycyl-(3S)-6,10-dithia-2-azaspiro[4.5]decane-3-carbonyl-3-amino-2-oxohexanoylglycyl-2-phenyl-glycinamide;

-   d)    N-acetyl-L-leucyl-(2S)-2-cyclohexylglycyl-(3S)-6,10-dithia-2-azaspiro[4.5]decane-3-carbonyl-3-amino-2-oxohexanoylglycyl-N,N-dimethyl-2-phenyl-glycinamide;

-   e)    N-acetyl-L-leucyl-(2S)-2-cyclohexylglycyl-(3S)-6,10-dithia-2-azaspiro[4.5]decane-3-carbonyl-3-amino-2-oxohexanoylglycyl-N-methoxy-N-methyl-2-phenyl-glycinamide;

-   f)    (2S)—N-acetyl-L-leucyl-(2S)-2-cyclohexylglycyl-(3S)-6,10-dithia-2-azaspiro[4.5]decane-3-carbonyl-3-amino-2-oxohexanoylglycyl-2-phenyl-1,1-dimethylethyl    ester, glycine;

-   g)    (2S)—N-acetyl-L-leucyl-(2S)-2-cyclohexylglycyl-(3S)-6,10-dithia-2-azaspiro[4.5]decane-3-carbonyl-3-amino-2-oxohexanoylglycyl-2-phenyl-glycine;

-   h)    (2S)—N-acetyl-L-leucyl-(2S)-2-cyclohexylglycyl-(3S)-6,10-dithia-2-azaspiro[4.5]decane-3-carbonyl-3-amino-2-oxohexanoylglycyl-2-phenyl-glycinamide;

-   i)    (2S)—N-acetyl-L-leucyl-(2S)-2-cyclohexylglycyl-(3S)-6,10-dithia-2-azaspiro[4.5]decane-3-carbonyl-3-amino-2-oxohexanoylglycyl-N,N-dimethyl-2-phenyl-glycinamide;

-   j)    (2S)—N-acetyl-L-leucyl-(2S)-2-cyclohexylglycyl-(3S)-6,10-dithia-2-azaspiro[4.5]decane-3-carbonyl-3-amino-2-oxohexanoylglycyl-N-methoxy-N-methyl-2-phenyl-glycinamide;

-   k)    N-acetyl-L-α-glutamyl-L-α-glutamyl-L-valyl-L-valyl-N-[1-[oxo(2-propenylamino)acetyl]butyl]-,bis(1,1-dimethylethyl)ester-(8S)-1,4-dithia-7-azaspiro[4.4]nonane-8-carboxamide;

-   l)    N-acetyl-L-α-glutamyl-L-α-glutamyl-L-valyl-L-valyl-N-[1-[oxo(2-propenylamino)acetyl]butyl]-,2-(1,1-dimethylethyl)ester-(8S)-1,4-dithia-7-azaspiro[4.4]nonane-8-carboxamide;

-   m)    N-acetyl-L-α-glutamyl-L-α-glutamyl-L-valyl-L-valyl-N-[1-[oxo(2-propenylamino)acetyl]butyl]-(8S)-1,4-dithia-7-azaspiro[4.4]nonane-8-carboxamide;

-   n)    N-acetyl-L-α-glutamyl-L-α-glutamyl-L-valyl-2-cyclohexylglycyl-(8S)-1,4-dithia-7-azaspiro[4.4]nonane-8-carbonyl-3-amino-2-oxohexanoyl-glycine;

-   o)    N-acetyl-L-α-glutamyl-L-α-glutamyl-L-valyl-2-cyclohexylglycyl-(8S)-1,4-dithia-7-azaspiro[4.4]nonane-8-carbonyl-3-amino-2-oxohexanoylglycyl-2-phenyl-glycinamide;

-   p)    N-acetyl-L-α-glutamyl-L-α-glutamyl-L-valyl-2-cyclohexylglycyl-(8S)-1,4-dithia-7-azaspiro[4.4]nonane-8-carbonyl-3-amino-2-oxohexanoyl-,1,2-bis(1,1-dimethylethyl)-7-(2-propenyl)ester    glycine; and

-   q)    N-acetyl-L-α-glutamyl-L-α-glutamyl-L-valyl-2-cyclohexylglycyl-(8S)-1,4-dithia-7-azaspiro[4.4]nonane-8-carbonyl-3-amino-2-oxohexanoyl-1,2-bis(1,1-dimethylethyl)ester    glycine.

The invention also relates to processes for preparing the abovecompounds and to compositions that comprise the above compounds and theuse thereof. Such compositions may be used to pre-treat invasive devicesto be inserted into a patient, to treat biological samples, such asblood, prior to administration to a patient, and for directadministration to a patient. In each case the composition will be usedto inhibit HCV replication and to lessen the risk of or the severity ofHCV infection.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides a compound of formula I:

or a pharmaceutically acceptable salt thereof,wherein:

-   X and X′ are both fluorine; or-   X and X′ are independently C(H), N, NH, O, or S; and X and X′ are    taken together with the carbon atom to which they are bound to form    a 5- to 7-membered saturated or partially unsaturated ring having up    to 4 heteroatoms independently selected from N, NH, O, S, SO, and    SO₂; wherein any atom is optionally singly or multiply substituted    with up to 3 substituents selected independently from J; and wherein    said ring is optionally fused to a second ring selected from    (C6-C10)aryl, (C5-C10)heteroaryl, (C3-C10)cycloalkyl, and a    (C3-C10)heterocyclyl, wherein said second ring has up to 3    substituents selected independently from J;    -   J is halogen, —OR′, —NO₂, —CN, —CF₃, —OCF₃, —R′, oxo, thioxo,        ═N(R′), ═N(OR′), 1,2-methylenedioxy, 1,2-ethylenedioxy, —N(R′)₂,        —SR′, —SOR′, —SO₂R′, —SO₂N(R′)₂, —SO₃R′, —C(O)R′, —C(O)C(O)R′,        —C(O)C(O)OR′, —C(O)C(O)NR′, —C(O)CH₂C(O)R′, —C(S)R′, —C(S)OR′,        —C(O)OR′, —OC(O)R′, —C(O)N(R′)₂, —OC(O)N(R′)₂, —C(S)N(R′)₂,        —(CH₂)₀₋₂NHC(O)R′, —N(R′)N(R′)COR′, —N(R′)N(R′)C(O)OR′,        —N(R′)N(R′)CON(R′)₂, —N(R′)SO₂R′, —N(R′)SO₂N(R′)₂,        —N(R′)C(O)OR′, —N(R′)C(O)R′, —N(R′)C(S)R′, —N(R′)C(O)N(R′)₂,        —N(R′)C(S)N(R′)₂, —N(COR′)COR′, —N(OR′)R′, —C(═NH)N(R′)₂,        —C(O)N(OR′)R′, —C(═NOR′)R′, —OP(O)(OR′)₂, —P(O)(R′)₂,        —P(O)(OR′)₂, or —P(O)(H)(OR′); wherein;        -   R′ is independently selected from:        -   hydrogen-,        -   (C1-C12)-aliphatic-,        -   (C3-C10)-cycloalkyl- or -cycloalkenyl-,        -   [(C3-C10)-cycloalkyl or -cycloalkenyl]-(C1-C12)-aliphatic-,        -   (C6-C10)-aryl-,        -   (C6-C10)-aryl-(C1-C12)aliphatic-,        -   (C3-C10)-heterocyclyl-,        -   (C3-C10)-heterocyclyl-(C1-C12)aliphatic-,        -   (C5-C10)-heteroaryl-, and        -   (C5-C10)-heteroaryl-(C1-C12)-aliphatic-;        -   wherein up to 5 atoms in R′ are optionally and independently            substituted with J;        -   wherein two R′ groups bound to the same atom form a 3- to            10-membered aromatic or non-aromatic ring having up to 3            heteroatoms independently selected from N, NH, O, S, SO, and            SO₂, wherein said ring is optionally fused to a            (C6-C10)aryl, (C5-C10)heteroaryl, (C3-C10)cycloalkyl, or a            (C3-C10)heterocyclyl, wherein any ring has up to 3            substituents selected independently from J;-   Y and Y′ are independently:    -   hydrogen-,    -   (C1-C12)-aliphatic-,    -   (C3-C10)-cycloalkyl- or -cycloalkenyl-,    -   (C3-C10)-cycloalkyl-(C1-C12)-aliphatic-,    -   (C6-C10)-aryl-,    -   (C3-C10)-heterocyclyl-; or    -   (C5-C10)-heteroaryl-;        -   wherein up to three aliphatic carbon atoms in Y and Y′ may            be replaced by O, N, NH, S, SO, or SO₂;        -   wherein each of Y and Y′ is independently and optionally            substituted with up to 3 substituents independently selected            from J;-   R₁ and R₃ are independently:    -   (C1-C12)-aliphatic-,    -   (C3-C10)-cycloalkyl- or -cycloalkenyl-,    -   [(C3-C10)-cycloalkyl- or -cycloalkenyl]-(C1-C12)-aliphatic-,    -   (C6-C10)-aryl-(C1-C12)aliphatic-, or    -   (C5-C10)-heteroaryl-(C1-C12)-aliphatic-;        -   wherein up to 3 aliphatic carbon atoms in R₁ and R₃ may be            replaced by a heteroatom selected from O, N, NH, S, SO, or            SO₂ in a chemically stable arrangement;        -   wherein each of R₁ and R₃ is independently and optionally            substituted with up to 3 substituents independently selected            from J;-   R₂, R₄, and R₇ are independently:    -   hydrogen-,    -   (C1-C12)-aliphatic-,    -   (C3-C10)-cycloalkyl-(C1-C12)-aliphatic-, or    -   (C6-C10)-aryl-(C1-C12)-aliphatic-;        -   wherein up to two aliphatic carbon atoms in R₂, R₄, and R₇            may be replaced by a heteroatom selected from O, N, NH, S,            SO, and SO₂ in a chemically stable arrangement;        -   wherein each of R₂, R₄, and R₇ is independently and            optionally substituted with up to 3 substituents            independently selected from J;-   R₅ and R_(5′) are independently hydrogen or (C1-C12)-aliphatic,    wherein any hydrogen is optionally replaced with halogen; wherein    any terminal carbon atom of R₅ is optionally substituted with    sulfhydryl or hydroxy; or R₅ is Ph or —CH₂Ph and R_(5′) is H,    wherein said Ph or —CH₂Ph group is optionally substituted with up to    3 substituents independently selected from J; or-   R₅ and R_(5′) together with the atom to which they are bound is a 3-    to 6-membered saturated or partially unsaturated ring having up to 2    heteroatoms selected from N, NH, O, SO, and SO₂; wherein the ring    has up to 2 substituents selected independently from J;-   W is:

-   -   wherein each R₆ is independently:        -   hydrogen-,        -   (C1-C12)-aliphatic-,        -   (C6-C10)-aryl-,        -   (C6-C10)-aryl-(C1-C12)aliphatic-,        -   (C3-C10)-cycloalkyl- or cycloalkenyl-,        -   [(C3-C10)-cycloalkyl- or cycloalkenyl]-(C1-C12)-aliphatic-,        -   (C3-C10)-heterocyclyl-,        -   (C3-C10)-heterocyclyl-(C1-C12)-aliphatic-,        -   (C5-C10)-heteroaryl-, or        -   (C5-C10)-heteroaryl-(C1-C12)-aliphatic-, or        -   two R₆ groups, which are bound to the same nitrogen atom,            form together with that nitrogen atom, a            (C3-C10)-heterocyclic ring;        -   wherein R₆ is optionally substituted with up to 3 J            substituents;    -   wherein each R₈ is independently —OR′; or the R₈ groups together        with the boron atom, is a (C3-C10)-membered heterocyclic ring        having in addition to the boron up to 3 additional heteroatoms        selected from N, NH, O, SO, and SO₂;

-   V is O or a valence bond; and

-   T is:    -   (C1-C12)-aliphatic-;    -   (C6-C10)-aryl-,    -   (C6-C10)-aryl-(C1-C12)aliphatic-,    -   (C3-C10)-cycloalkyl or -cycloalkenyl-,    -   [(C3-C10)-cycloalkyl or -cycloalkenyl]-(C1-C12)-aliphatic-,    -   (C3-C10)-heterocyclyl-,    -   (C3-C10)-heterocyclyl-(C1-C12)-aliphatic-,    -   (C5-C10)-heteroaryl-, or    -   (C5-C10)-heteroaryl-(C1-C12)-aliphatic-;        -   wherein up to 3 aliphatic carbon atoms in T may be replaced            by a heteroatom selected from O, N, NH, S, SO, or SO₂ in a            chemically stable arrangement;        -   wherein each T is optionally substituted with up to 3 J            substituents;            provided that the following compounds are excluded:

-   a)    N-acetyl-L-leucyl-(2S)-2-cyclohexylglycyl-(3S)-6,10-dithia-2-azaspiro[4.5]decane-3-carbonyl-3-amino-2-oxohexanoylglycyl-2-phenyl-1,1-dimethylethyl    ester glycine;

-   b)    N-acetyl-L-leucyl-(2S)-2-cyclohexylglycyl-(3S)-6,10-dithia-2-azaspiro[4.5]decane-3-carbonyl-3-amino-2-oxohexanoylglycyl-2-phenyl-glycine;

-   c)    N-acetyl-L-leucyl-(2S)-2-cyclohexylglycyl-(3S)-6,10-dithia-2-azaspiro[4.5]decane-3-carbonyl-3-amino-2-oxohexanoylglycyl-2-phenyl-glycinamide;

-   d)    N-acetyl-L-leucyl-(2S)-2-cyclohexylglycyl-(3S)-6,10-dithia-2-azaspiro[4.5]decane-3-carbonyl-3-amino-2-oxohexanoylglycyl-N,N-dimethyl-2-phenyl-glycinamide;

-   e)    N-acetyl-L-leucyl-(2S)-2-cyclohexylglycyl-(3S)-6,10-dithia-2-azaspiro[4.5]decane-3-carbonyl-3-amino-2-oxohexanoylglycyl-N-methoxy-N-methyl-2-phenyl-glycinamide;

-   f)    (2S)—N-acetyl-L-leucyl-(2S)-2-cyclohexylglycyl-(3S)-6,10-dithia-2-azaspiro[4.5]decane-3-carbonyl-3-amino-2-oxohexanoylglycyl-2-phenyl-1,1-dimethylethyl    ester, glycine;

-   g)    (2S)—N-acetyl-L-leucyl-(2S)-2-cyclohexylglycyl-(3S)-6,10-dithia-2-azaspiro[4.5]decane-3-carbonyl-3-amino-2-oxohexanoylglycyl-2-phenyl-glycine;

-   h)    (2S)—N-acetyl-L-leucyl-(2S)-2-cyclohexylglycyl-(3S)-6,10-dithia-2-azaspiro[4.5]decane-3-carbonyl-3-amino-2-oxohexanoylglycyl-2-phenyl-glycinamide;

-   i)    (2S)—N-acetyl-L-leucyl-(2S)-2-cyclohexylglycyl-(3S)-6,10-dithia-2-azaspiro[4.5]decane-3-carbonyl-3-amino-2-oxohexanoylglycyl-N,N-dimethyl-2-phenyl-glycinamide;

-   j)    (2S)—N-acetyl-L-leucyl-(2S)-2-cyclohexylglycyl-(3S)-6,10-dithia-2-azaspiro[4.5]decane-3-carbonyl-3-amino-2-oxohexanoylglycyl-N-methoxy-N-methyl-2-phenyl-glycinamide;

-   k)    N-acetyl-L-α-glutamyl-L-α-glutamyl-L-valyl-L-valyl-N-[1-[oxo(2-propenylamino)acetyl]butyl]-,bis(1,1-dimethylethyl)ester-(8S)-1,4-dithia-7-azaspiro[4.4]nonane-8-carboxamide;

-   l)    N-acetyl-L-α-glutamyl-L-α-glutamyl-L-valyl-L-valyl-N-[1-[oxo(2-propenylamino)acetyl]butyl]-,2-(1,1-dimethylethyl)ester-(8S)-1,4-dithia-7-azaspiro[4.4]nonane-8-carboxamide;

-   m)    N-acetyl-L-α-glutamyl-L-α-glutamyl-L-valyl-L-valyl-N-[1-[oxo(2-propenylamino)acetyl]butyl]-(8S)-1,4-dithia-7-azaspiro[4.4]nonane-8-carboxamide;

-   n)    N-acetyl-L-α-glutamyl-L-α-glutamyl-L-valyl-2-cyclohexylglycyl-(8S)-1,4-dithia-7-azaspiro[4.4]nonane-8-carbonyl-3-amino-2-oxohexanoyl-glycine;

-   o)    N-acetyl-L-α-glutamyl-L-α-glutamyl-L-valyl-2-cyclohexylglycyl-(8S)-1,4-dithia-7-azaspiro[4.4]nonane-8-carbonyl-3-amino-2-oxohexanoylglycyl-2-phenyl-glycinamide;

-   p)    N-acetyl-L-α-glutamyl-L-α-glutamyl-L-valyl-2-cyclohexylglycyl-(8S)-1,4-dithia-7-azaspiro[4.4]nonane-8-carbonyl-3-amino-2-oxohexanoyl-,1,2-bis(1,1-dimethylethyl)-7-(2-propenyl)ester    glycine; and

-   q)    N-acetyl-L-α-glutamyl-L-α-glutamyl-L-valyl-2-cyclohexylglycyl-(8S)-1,4-dithia-7-azaspiro[4.4]nonane-8-carbonyl-3-amino-2-oxohexanoyl-1,2-bis(1,1-dimethylethyl)ester    glycine.

DEFINITIONS

The term “aryl” as used herein means a monocyclic or bicycliccarbocyclic aromatic ring system. Phenyl is an example of a monocyclicaromatic ring system. Bicyclic aromatic ring systems include systemswherein both rings are aromatic, e.g., naphthyl, and systems whereinonly one of the two rings is aromatic, e.g., tetralin. It is understoodthat as used herein, the term “(C6-C10)-aryl-” includes any one of a C6,C7, C8, C9, and C10 monocyclic or bicyclic carbocyclic aromatic ring.

The term “heterocyclyl” as used herein means a monocyclic or bicyclicnon-aromatic ring system having 1 to 3 heteroatom or heteroatom groupsin each ring selected from O, N, NH, S, SO, and SO₂ in a chemicallystable arrangement. In a bicyclic non-aromatic ring system embodiment of“heterocyclyl” one or both rings may contain said heteroatom orheteroatom groups. It is understood that as used herein, the term“(C5-C10)-heterocyclyl-” includes any one of a C5, C6, C7, C8, C9, andC10 monocyclic or bicyclic non-aromatic ring system having 1 to 3heteroatom or heteroatom groups in each ring selected from O, N, NH, andS in a chemically stable arrangement.

The term “heteroaryl” as used herein means a monocyclic or bicyclicaromatic ring system having 1 to 3 heteroatom or heteroatom groups ineach ring selected from O, N, NH, and S in a chemically stablearrangement. In such a bicyclic aromatic ring system embodiment of“heteroaryl”:

-   -   one or both rings may be aromatic; and    -   one or both rings may contain said heteroatom or heteroatom        groups. It is understood that as used herein, the term        “(C5-C10)-heteroaryl-” includes any one of a C5, C6, C7, C8, C9,        and C10 monocyclic or bicyclic aromatic ring system having 1 to        3 heteroatom or heteroatom groups in each ring selected from O,        N, NH, and S in a chemically stable arrangement.

The term “aliphatic” as used herein means a straight chained or branchedalkyl, alkenyl or alkynyl. It is understood that as used herein, theterm “(C1-C12)-aliphatic-” includes any one of a C1, C2, C3, C4, C5, C6,C7, C8, C9, C10, C11, and C12 straight or branched alkyl chain of carbonatoms. It is also understood that alkenyl or alkynyl embodiments need atleast two carbon atoms in the aliphatic chain. The term “cycloalkyl orcycloalkenyl” refers to a monocyclic or fused or bridged bicycliccarbocyclic ring system that is not aromatic. Cycloalkenyl rings haveone or more units of unsaturation. It is also understood that as usedherein, the term “(C3-C10)-cycloalkyl- or -cycloalkenyl-” includes anyone of a C3, C4, C5, C6, C7, C8, C9, and C10 monocyclic or fused orbridged bicyclic carbocyclic ring. Examples of cycloalkyl groups includecyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl,cycloheptyl, cycloheptenyl, nornbornyl, adamantyl and decalin-yl.

The phrase “chemically stable arrangement” as used herein refers to acompound structure that renders the compound sufficiently stable toallow manufacture and administration to a mammal by methods known in theart. Typically, such compounds are stable at a temperature of 40° C. orless, in the absence of moisture or other chemically reactive condition,for at least a week.

EMBODIMENTS

According to one embodiment, the compounds of the present invention areof formula I:

or a pharmaceutically acceptable salt thereof,wherein:

-   X and X′ are both fluorine; or-   X and X′ are independently C(H), N, NH, O, or S; and X and X′ are    taken together with the carbon atom to which they are bound to form    a 5- to 7-membered saturated or partially unsaturated ring having up    to 4 heteroatoms independently selected from N, NH, O, S, SO, and    SO₂; wherein any atom is optionally singly or multiply substituted    with up to 3 substituents selected independently from J; and wherein    said ring is optionally fused to a second ring selected from    (C6-C10)aryl, (C5-C10)heteroaryl, (C3-C10)cycloalkyl, and a    (C3-C10)heterocyclyl, wherein said second ring has up to 3    substituents selected independently from J;    -   J is halogen, —OR′, —NO₂, —CN, —CF₃, —OCF₃, —R′, oxo, thioxo,        ═N(R′), ═N(OR′), 1,2-methylenedioxy, 1,2-ethylenedioxy, —N(R′)₂,        —SR′, —SOR′, —SO₂R′, —SO₂N(R′)₂, —SO₃R′, —C(O)R′, —C(O)C(O)R′,        —C(O)C(O)OR′, —C(O)C(O)NR′, —C(O)CH₂C(O)R′, —C(S)R′, —C(S)OR′,        —C(O)OR′, —OC(O)R′, —C(O)N(R′)₂, —OC(O)N(R′)₂, —C(S)N(R′)₂,        —(CH₂)₀₋₂NHC(O)R′, —N(R′)N(R′)COR′, —N(R′)N(R′)C(O)OR′,        —N(R′)N(R′)CON(R′)₂, —N(R′)SO₂R′, —N(R′)SO₂N(R′)₂,        —N(R′)C(O)OR′, —N(R′)C(O)R′, —N(R′)C(S)R′, —N(R′)C(O)N(R′)₂,        —N(R′)C(S)N(R′)₂, —N(COR′)COR′, —N(OR′)R′, —C(═NH)N(R′)₂,        —C(O)N(OR′)R′, —C(═NOR′)R′, —OP(O)(OR′)₂, —P(O)(R′)₂,        —P(O)(OR′)₂, or —P(O)(H)(OR′); wherein;        -   R′ is independently selected from:        -   hydrogen-,        -   (C1-C12)-aliphatic-,        -   (C3-C10)-cycloalkyl- or -cycloalkenyl-,        -   [(C3-C10)-cycloalkyl or -cycloalkenyl]-(C1-C12)-aliphatic-,        -   (C6-C10)-aryl-,        -   (C6-C10)-aryl-(C1-C12)aliphatic-,        -   (C3-C10)-heterocyclyl-,        -   (C3-C10)-heterocyclyl-(C1-C12)aliphatic-,        -   (C5-C10)-heteroaryl-, and        -   (C5-C10)-heteroaryl-(C1-C12)-aliphatic-;        -   wherein up to 5 atoms in R′ are optionally and independently            substituted with J;        -   wherein two R′ groups bound to the same atom form a 3- to            10-membered aromatic or non-aromatic ring having up to 3            heteroatoms independently selected from N, NH, O, S, SO, and            SO₂, wherein said ring is optionally fused to a            (C6-C10)aryl, (C5-C10)heteroaryl, (C3-C10)cycloalkyl, or a            (C3-C10)heterocyclyl, wherein any ring has up to 3            substituents selected independently from J;-   Y and Y′ are independently:    -   hydrogen-,    -   (C1-C12)-aliphatic-,    -   (C3-C10)-cycloalkyl- or -cycloalkenyl-,    -   (C3-C10)-cycloalkyl-(C1-C12)-aliphatic-,    -   (C6-C10)-aryl-,    -   (C3-C10)-heterocyclyl-; or    -   (C5-C10)-heteroaryl-;        -   wherein up to three aliphatic carbon atoms in Y and Y′ may            be replaced by O, N, NH, S, SO, or SO₂;        -   wherein each of Y and Y′ is independently and optionally            substituted with up to 3 substituents independently selected            from J;-   R₁ and R₃ are independently:    -   (C1-C12)-aliphatic-,    -   (C3-C10)-cycloalkyl- or -cycloalkenyl-,    -   [(C3-C10)-cycloalkyl- or -cycloalkenyl]-(C1-C12)-aliphatic-,    -   (C6-C10)-aryl-(C1-C12)aliphatic-, or    -   (C5-C10)-heteroaryl-(C1-C12)-aliphatic-;        -   wherein up to 3 aliphatic carbon atoms in R₁ and R₃ may be            replaced by a heteroatom selected from O, N, NH, S, SO, or            SO₂ in a chemically stable arrangement;        -   wherein each of R₁ and R₃ is independently and optionally            substituted with up to 3 substituents independently selected            from J;-   R₂, R₄, and R₇ are independently:    -   hydrogen-,    -   (C1-C12)-aliphatic-,    -   (C3-C10)-cycloalkyl-(C1-C12)-aliphatic-, or    -   (C6-C10)-aryl-(C1-C12)-aliphatic-;        -   wherein up to two aliphatic carbon atoms in R₂, R₄, and R₇            may be replaced by a heteroatom selected from O, N, NH, S,            SO, and SO₂ in a chemically stable arrangement;        -   wherein each of R₂, R₄, and R₇ is independently and            optionally substituted with up to 3 substituents            independently selected from J;-   R₅ and R_(5′) are independently hydrogen or (C1-C12)-aliphatic,    wherein any hydrogen is optionally replaced with halogen; wherein    any terminal carbon atom of R₅ is optionally substituted with    sulfhydryl or hydroxy; or R₅ is Ph or —CH₂Ph and R_(5′) is H,    wherein said Ph or —CH₂Ph group is optionally substituted with up to    3 substituents independently selected from J; or-   R₅ and R_(5′) together with the atom to which they are bound is a 3-    to 6-membered saturated or partially unsaturated ring having up to 2    heteroatoms selected from N, NH, O, SO, and SO₂; wherein the ring    has up to 2 substituents selected independently from J;-   W is:

-   -   wherein each R₆ is independently:        -   hydrogen-,        -   (C1-C12)-aliphatic-,        -   (C6-C10)-aryl-,        -   (C6-C10)-aryl-(C1-C12)aliphatic-,        -   (C3-C10)-cycloalkyl- or cycloalkenyl-,        -   [(C3-C10)-cycloalkyl- or cycloalkenyl]-(C1-C12)-aliphatic-,        -   (C3-C10)-heterocyclyl-,        -   (C3-C10)-heterocyclyl-(C1-C12)-aliphatic-,        -   (C5-C10)-heteroaryl-, or        -   (C5-C10)-heteroaryl-(C1-C12)-aliphatic-, or        -   two R₆ groups, which are bound to the same nitrogen atom,            form together with that nitrogen atom, a            (C3-C10)-heterocyclic ring;        -   wherein R₆ is optionally substituted with up to 3 J            substituents;    -   wherein each R₈ is independently —OR′; or the R₈ groups together        with the boron atom, is a (C3-C10)-membered heterocyclic ring        having in addition to the boron up to 3 additional heteroatoms        selected from N, NH, O, SO, and SO₂;

-   V is O or a valence bond; and

-   T is:    -   (C1-C12)-aliphatic-;    -   (C6-C10)-aryl-,    -   (C6-C10)-aryl-(C1-C12)aliphatic-,    -   (C3-C10)-cycloalkyl or -cycloalkenyl-,    -   [(C3-C10)-cycloalkyl or -cycloalkenyl]-(C1-C12)-aliphatic-,    -   (C3-C10)-heterocyclyl-,    -   (C3-C10)-heterocyclyl-(C1-C12)-aliphatic-,    -   (C5-C10)-heteroaryl-, or    -   (C5-C10)-heteroaryl-(C1-C12)-aliphatic-;        -   wherein up to 3 aliphatic carbon atoms in T may be replaced            by a heteroatom selected from O, N, NH, S, SO, or SO₂ in a            chemically stable arrangement;        -   wherein each T is optionally substituted with up to 3 J            substituents;            provided that the following compounds are excluded:

-   a)    N-acetyl-L-leucyl-(2S)-2-cyclohexylglycyl-(3S)-6,10-dithia-2-azaspiro[4.5]decane-3-carbonyl-3-amino-2-oxohexanoylglycyl-2-phenyl-1,1-dimethylethyl    ester glycine;

-   b)    N-acetyl-L-leucyl-(2S)-2-cyclohexylglycyl-(3S)-6,10-dithia-2-azaspiro[4.5]decane-3-carbonyl-3-amino-2-oxohexanoylglycyl-2-phenyl-glycine;

-   c)    N-acetyl-L-leucyl-(2S)-2-cyclohexylglycyl-(3S)-6,10-dithia-2-azaspiro[4.5]decane-3-carbonyl-3-amino-2-oxohexanoylglycyl-2-phenyl-glycinamide;

-   d)    N-acetyl-L-leucyl-(2S)-2-cyclohexylglycyl-(3S)-6,10-dithia-2-azaspiro[4.5]decane-3-carbonyl-3-amino-2-oxohexanoylglycyl-N,N-dimethyl-2-phenyl-glycinamide;

-   e)    N-acetyl-L-leucyl-(2S)-2-cyclohexylglycyl-(3S)-6,10-dithia-2-azaspiro[4.5]decane-3-carbonyl-3-amino-2-oxohexanoylglycyl-N-methoxy-N-methyl-2-phenyl-glycinamide;

-   f)    (2S)—N-acetyl-L-leucyl-(2S)-2-cyclohexylglycyl-(3S)-6,10-dithia-2-azaspiro[4.5]decane-3-carbonyl-3-amino-2-oxohexanoylglycyl-2-phenyl-1,1-dimethylethyl    ester, glycine;

-   g)    (2S)—N-acetyl-L-leucyl-(2S)-2-cyclohexylglycyl-(3S)-6,10-dithia-2-azaspiro[4.5]decane-3-carbonyl-3-amino-2-oxohexanoylglycyl-2-phenyl-glycine;

-   h)    (2S)—N-acetyl-L-leucyl-(2S)-2-cyclohexylglycyl-(3S)-6,10-dithia-2-azaspiro[4.5]decane-3-carbonyl-3-amino-2-oxohexanoylglycyl-2-phenyl-glycinamide;

-   i)    (2S)—N-acetyl-L-leucyl-(2S)-2-cyclohexylglycyl-(3S)-6,10-dithia-2-azaspiro[4.5]decane-3-carbonyl-3-amino-2-oxohexanoylglycyl-N,N-dimethyl-2-phenyl-glycinamide;

-   j)    (2S)—N-acetyl-L-leucyl-(2S)-2-cyclohexylglycyl-(3S)-6,10-dithia-2-azaspiro[4.5]decane-3-carbonyl-3-amino-2-oxohexanoylglycyl-N-methoxy-N-methyl-2-phenyl-glycinamide;

-   k)    N-acetyl-L-α-glutamyl-L-α-glutamyl-L-valyl-L-valyl-N-[1-[oxo(2-propenylamino)acetyl]butyl]-,bis(1,1-dimethylethyl)ester-(8S)-1,4-dithia-7-azaspiro[4.4]nonane-8-carboxamide;

-   l)    N-acetyl-L-α-glutamyl-L-α-glutamyl-L-valyl-L-valyl-N-[1-[oxo(2-propenylamino)acetyl]butyl]-,2-(1,1-dimethylethyl)ester-(8S)-1,4-dithia-7-azaspiro[4.4]nonane-8-carboxamide;

-   m)    N-acetyl-L-α-glutamyl-L-α-glutamyl-L-valyl-L-valyl-N-[1-[oxo(2-propenylamino)acetyl]butyl]-(8S)-1,4-dithia-7-azaspiro[4.4]nonane-8-carboxamide;

-   n)    N-acetyl-L-α-glutamyl-L-α-glutamyl-L-valyl-2-cyclohexylglycyl-(8S)-1,4-dithia-7-azaspiro[4.4]nonane-8-carbonyl-3-amino-2-oxohexanoyl-glycine;

-   o)    N-acetyl-L-α-glutamyl-L-α-glutamyl-L-valyl-2-cyclohexylglycyl-(8S)-1,4-dithia-7-azaspiro[4.4]nonane-8-carbonyl-3-amino-2-oxohexanoylglycyl-2-phenyl-glycinamide;

-   p)    N-acetyl-L-α-glutamyl-L-α-glutamyl-L-valyl-2-cyclohexylglycyl-(8S)-1,4-dithia-7-azaspiro[4.4]nonane-8-carbonyl-3-amino-2-oxohexanoyl-,1,2-bis(1,1-dimethylethyl)-7-(2-propenyl)ester    glycine; and

-   q)    N-acetyl-L-α-glutamyl-L-α-glutamyl-L-valyl-2-cyclohexylglycyl-(8S)-1,4-dithia-7-azaspiro[4.4]nonane-8-carbonyl-3-amino-2-oxohexanoyl-1,2-bis(1,1-dimethylethyl)ester    glycine.

According to another embodiment of the present invention, the compoundsare of formula Ia:

or a pharmaceutically acceptable salt thereof,wherein:

-   X and X′ are independently C(H), N, NH, O, or S; and X and X′ are    taken together with the carbon atom to which they are bound to form    a 5- to 7-membered saturated or partially unsaturated spirocyclic    ring having up to 4 heteroatoms independently selected from N, NH,    O, S, SO, and SO₂; wherein any atom is optionally singly or multiply    substituted with up to 3 substituents selected independently from J;    and wherein said ring is optionally fused to a second ring selected    from (C6-C10)aryl, (C5-C10)heteroaryl, (C3-C10)cycloalkyl, and a    (C3-C10)heterocyclyl, wherein said second ring has up to 3    substituents selected independently from J;    -   J is halogen, —OR′, —NO₂, —CN, —CF₃, —OCF₃, —R′, oxo, thioxo,        ═N(R′), ═N(OR′), 1,2-methylenedioxy, 1,2-ethylenedioxy, —N(R′)₂,        —SR′, —SOR′, —SO₂R′, —SO₂N(R′)₂, —SO₃R′, —C(O)R′, —C(O)C(O)R′,        —C(O)C(O)OR′, —C(O)C(O)NR′, —C(O)CH₂C(O)R′, —C(S)R′, —C(S)OR′,        —C(O)OR′, —OC(O)R′, —C(O)N(R′)₂, —OC(O)N(R′)₂, —C(S)N(R′)₂,        —(CH₂)₀₋₂NHC(O)R′, —N(R′)N(R′)COR′, —N(R′)N(R′)C(O)OR′,        —N(R′)N(R′)CON(R′)₂, —N(R′)SO₂R′, —N(R′)SO₂N(R′)₂,        —N(R′)C(O)OR′, —N(R′)C(O)R′, —N(R′)C(S)R′, —N(R′)C(O)N(R′)₂,        —N(R′)C(S)N(R′)₂, —N(COR′)COR′, —N(OR′)R′, —C(═NH)N(R′)₂,        —C(O)N(OR′)R′, —C(═NOR′)R′, —OP(O)(OR′)₂, —P(O)(R′)₂,        —P(O)(OR′)₂, or —P(O)(H)(OR′); wherein;        -   R′ is independently selected from:        -   hydrogen-,        -   (C1-C12)-aliphatic-,        -   (C3-C10)-cycloalkyl- or -cycloalkenyl-,        -   [(C3-C10)-cycloalkyl or -cycloalkenyl]-(C1-C12)-aliphatic-,        -   (C6-C10)-aryl-,        -   (C6-C10)-aryl-(C1-C12)aliphatic-,        -   (C3-C10)-heterocyclyl-,        -   (C3-C10)-heterocyclyl-(C1-C12)aliphatic-,        -   (C5-C10)-heteroaryl-, and        -   (C5-C10)-heteroaryl-(C1-C12)-aliphatic-;        -   wherein up to 5 atoms in R′ are optionally and independently            substituted with J;        -   wherein two R′ groups bound to the same atom form a 3- to            10-membered aromatic or non-aromatic ring having up to 3            heteroatoms independently selected from N, NH, O, S, SO, and            SO₂, wherein said ring is optionally fused to a            (C6-C10)aryl, (C5-C10)heteroaryl, (C3-C10)cycloalkyl, or a            (C3-C10)heterocyclyl, wherein any ring has up to 3            substituents selected independently from J;-   Y and Y′ are hydrogen;-   R₁ and R₃ are independently:    -   (C1-C6)-aliphatic-,    -   (C3-C10)-cycloalkyl- or -cycloalkenyl-,    -   [(C3-C10)-cycloalkyl- or -cycloalkenyl]-(C1-C6)-aliphatic-, or    -   (C6-C10)-aryl-(C1-C6)aliphatic-;        -   wherein up to 3 aliphatic carbon atoms in R₁ and R₃ may be            replaced by a heteroatom selected from O, N, NH, S, SO, or            SO₂ in a chemically stable arrangement;        -   wherein each of R₁ and R₃ is independently and optionally            substituted with up to 3 substituents independently selected            from J;-   R₂ and R₇ are hydrogen;-   R₄ is selected from:    -   hydrogen-,    -   (C1-C6)-alkyl-,    -   (C3-C10)-cycloalkyl-(C1-C6)-aliphatic-, or    -   (C6-C10)-aryl-(C1-C6)-aliphatic-;        -   wherein R₄ is independently and optionally substituted with            up to 3 substituents independently selected from J;    -   R_(5′) is hydrogen;-   R₅ is (C1-C6)-aliphatic, wherein any hydrogen is optionally replaced    with halogen;-   W is:

-   -   wherein each R₆ is independently:        -   hydrogen-,        -   (C1-C6)-alkyl-,        -   (C6-C10)-aryl-,        -   (C6-C10)-aryl-(C1-C6)alkyl-,        -   (C3-C10)-cycloalkyl- or cycloalkenyl-,        -   [(C3-C10)-cycloalkyl- or cycloalkenyl]-(C1-C6)-alkyl-,        -   (C5-C10)-heteroaryl-(C1-C6)-alkyl-, or        -   two R₆ groups, which are bound to the same nitrogen atom,            form together with that nitrogen atom, a            (C3-C10)-heterocyclic ring;        -   wherein R₆ is optionally substituted with up to 3 J            substituents;        -   wherein each R₈ is independently —OR′; or the R₈ groups            together with the boron atom, is a (C3-C10)-membered            heterocyclic ring having in addition to the boron up to 3            additional heteroatoms selected from N, NH, O, SO, and SO₂;

-   V is a valence bond; and

-   T is:    -   (C6-C10)-aryl-,    -   (C6-C10)-aryl-(C1-C6)aliphatic-,    -   (C5-C10)-heteroaryl-, or    -   (C5-C10)-heteroaryl-(C1-C6)-aliphatic-;        -   wherein up to 3 aliphatic carbon atoms in T may be replaced            by a heteroatom selected from O, N, NH, S, SO, or SO₂ in a            chemically stable arrangement;        -   wherein each T is optionally substituted with up to 3 J            substituents.

According to one embodiment of compounds of formula I, the

radical is:

wherein:

-   -   n is 0, 1, or 2;    -   Y and Y′ are as defined in any of the embodiments herein; and    -   the ring containing X and X′ is optionally substituted with up        to 3 J substituents, wherein J is as defined in any of the        embodiments herein.

According to another embodiment for compounds of formula I, the

radical is:

wherein:

-   -   n is 0, 1, or 2;    -   Y and Y′ are as defined above; and the ring containing X and X′        is optionally substituted with up to 3 J substituents, wherein J        is as defined in any of the embodiments herein.

According to another embodiment for compounds of formula I, the

radical is:

wherein:

-   -   n is 0 or 1; and    -   Y and Y′ are H.

In another embodiment of compounds of formula I, the

radical is:

wherein:

-   -   n is 0 or 1; and    -   Y and Y′ are H.

In another embodiment of compounds of formula I, the

radical is:

wherein:

-   -   n is 1; and    -   Y and Y′ are H.

According to one embodiment of compounds of formula Ia, the

radical is:

wherein:

-   -   n is 0, 1, or 2; and    -   the ring containing X and X′ is optionally substituted with up        to 3 J substituents, wherein J is as defined in any of the        embodiments herein.

According to another embodiment of compounds of formula Ia, the

radical is:

wherein:

-   -   n is 0, 1, or 2; and    -   the ring containing X and X′ is optionally substituted with up        to 3 J substituents, wherein J is as defined in any of the        embodiments herein.

According to another embodiment of compounds of formula Ia, the

radical is:

wherein:

-   -   n is 0 or 1.

According to another embodiment of compounds of formula Ia, the

radical is:

wherein:

-   -   n is 0 or 1.

According to another embodiment of compounds of formula I or formula Ia,the present invention provides a compound of formula ID:

wherein:

-   -   n, R₁, R₂, R₃, R₄, R₅, R_(5′), R₇, V, T, W, Y, and Y′ are as        defined in any of the embodiments herein.

According to an embodiment of compounds of formula I, the

radical is:

wherein:

-   -   X, X′, Y, and Y′ are as defined in any of the embodiments        herein; and    -   the fused benzo ring is optionally substituted with up to 3 J        substituents, wherein J is as defined in any of the embodiments        herein.

According to another embodiment of compounds of formula I, the

radical is:

wherein:

-   -   X and X′, are as defined in any of the embodiments herein; Y and        Y′ are H; and    -   the fused benzo ring is optionally substituted with up to 3 J        substituents, wherein J is as defined in any of the embodiments        herein.

According to an embodiment of compounds of formula Ia, the

radical is:

wherein:

-   -   X, and X′, are as defined in any of the embodiments herein; and    -   the fused benzo ring is optionally substituted with up to 3 J        substituents, wherein J is as defined in any of the embodiments        herein.

According to another embodiment of compounds of formula Ia, the

radical is:

wherein:

-   -   X and X′, are as defined in any of the embodiments herein; Y and        Y′ are H; and    -   the fused benzo ring is optionally substituted with up to 3 J        substituents, wherein J is as defined in any of the embodiments        herein.

According to another embodiment of compounds of formula I or formula Ia,the present invention provides a compound of formula IE:

wherein:

-   -   R₁, R₂, R₃, R₄, R₅, R_(5′), R₇, V, T, W, Y, Y′, X and X′ are as        defined in any of the embodiments herein; and    -   the fused benzo ring is optionally substituted with up to 3 J        substituents, wherein J is as defined in any of the embodiments        herein.

According to another embodiment for compounds of formula IE, X and X′are S, Y and Y′ are H, R₁, R₂, R₃, R₄, R₅, R_(5′), R₇, V, T, and W areas defined in any of the embodiments herein, and the fused benzo ring isoptionally substituted with up to 3 J substituents, wherein J is asdefined in any of the embodiments herein.

According to another embodiment for compounds of formula I, the

radical is:

wherein:

-   -   Y and Y′ are H.

In another embodiment of compounds of formula I, the

radical is:

wherein:

-   -   Y and Y′ are H.

According to yet another embodiment of compounds of formula I, thepresent invention provides a compound of formula IF:

wherein:

-   -   R₁, R₂, R₃, R₄, R₅, R_(5′), R₇, V, T, W, Y and Y′ are as defined        in any of the embodiments herein.

According to another embodiment of compounds of formula I, W is:

wherein in the W, the NR₆R₆ is selected from —NH—(C1-C6 aliphatic),—NH—(C3-C6 cycloalkyl), —NH—CH(CH₃)-aryl, or —NH—CH(CH₃)-heteroaryl,wherein said aryl or said heteroaryl is optionally substituted with upto 3 halogens.

According to another embodiment of compounds of formula I, W is:

wherein in the W, when the NR₆R₆ is selected from —NH—(C1-C6 aliphatic),said C1-C6 aliphatic is C1-C6 alkyl with no J substituents.

According to another embodiment of compounds of formula I, W is:

wherein in the W, when the NR₆R₆ is selected from either(C6-C10)-aryl-(C1-C12)-aliphatic or(C5-C10)-heteroaryl-(C1-C12)-aliphatic-, said C1-C12-aliphatic is aC1-C6 alkyl group with no J substituents. In another embodiment, saidC1-C6 alkyl is substituted with up to 3 J substituents.

According to another embodiment in compounds of formula I, the NR₆R₆ inthe W radical is:

According to another embodiment in compounds of formula I, the NR₆R₆ inthe W radical is:

In another embodiment of compounds of formula I, in the W, the NR₆R₆ is:

In yet another embodiment of compounds of formula I, in the W, the NR₆R₆is:

According to an embodiment in compounds of formula I or formula Ia, theNR₆R₆ in the W radical is:

According to another embodiment in compounds of formula I or formula Ia,the NR₆R₆ in the W radical is:

In another embodiment of compounds of formula I or formula Ia, in the W,the NR₆R₆ is:

According to another embodiment of compounds of formula I, W is:

wherein in the W, the NR₆R₆ is NH₂.

According to another embodiment of compounds of formula I, W is:

wherein in the W, the R₆ is as defined in any of the embodiments herein.

According to another embodiment of compounds of formula I, W is:

wherein in the W, the R₆ is as defined in any of the embodiments herein.

According to another embodiment of compounds of formula I, W is:

wherein in the W, the R₈ is as defined in any of the embodiments herein.

According to an embodiment of compounds of formula Ia, W is:

wherein in the W, the NR₆R₆ is selected from —NH—(C1-C6 aliphatic),—NH—(C3-C6 cycloalkyl), —NH—CH(CH₃)-aryl, or —NH—CH(CH₃)-heteroaryl,wherein said aryl or said heteroaryl is optionally substituted with upto 3 halogens.

According to another embodiment of compounds of formula Ia, W is:

wherein in the W, when the NR₆R₆ is selected from —NH—(C1-C6 alkyl),said C1-C6 alkyl has no J substituents.

According to another embodiment of compounds of formula Ia, W is:

wherein in the W, when the NR₆R₆ is selected from either(C6-C10)-aryl-(C1-C6)-alkyl or (C5-C10)-heteroaryl-(C1-C6)-alkyl-, saidC1-C6 alkyl group has no J substituents. In another embodiment, saidC1-C6 alkyl is substituted with up to 3 J substituents.

According to another embodiment in compounds of formula Ia, the NR₆R₆ inthe W radical is:

According to another embodiment in compounds of formula Ia, the NR₆R₆ inthe W radical is:

In another embodiment of compounds of formula Ia, in the W, the NR₆R₆is:

In yet another embodiment of compounds of formula Ia, in the W, theNR6R₆ is:

According to another embodiment of compounds of formula Ia, W is:

wherein in the W, the NR₆R₆ is NH₂.

According to another embodiment of compounds of formula Ia, W is:

wherein in the W, the R₆ is as defined in any of the embodiments herein.

According to another embodiment of compounds of formula Ia, W is:

wherein in the W, the R₆ is as defined in any of the embodiments herein.

According to another embodiment of compounds of formula Ia, W is:

wherein in the W, the R₈ is as defined in any of the embodiments herein.

According to another embodiment for W in compounds of formula I orformula Ia, each R₈ together with the boron atom, is a (C5-C10)-memberedheterocyclic ring having no additional heteroatoms other than the boronand the two oxygen atoms. In one embodiment, groups are selected from:

wherein R′ is (C1-C6)-aliphatic. In another embodiment of compounds offormula I or formula IA, R′ is methyl.

According to yet another embodiment of compounds of formula I, thepresent invention provides a compound of formula IG:

wherein:

n, R₁, R₂, R₃, R₄, R₅, R_(5′), R₇, V, T, X, X′, Y, and Y′ are as definedin any of the embodiments herein.

According to another embodiment for compounds of formula IG, X and X′are S, Y and Y′ are H, n, R₁, R₂, R₃, R₄, R₅, R_(5′), R₇, V, and T areas defined in any of the embodiments herein.

According to another embodiment of compounds of formula I or formula Ia,the present invention provides a compound of formula IG-1:

wherein:

-   -   n, R₁, R₂, R₃, R₄, R₅, R_(5′), R₇, V, T, X, X′, Y, and Y′ are as        defined in any of the embodiments herein.

According to another embodiment of compounds of formula IG-1, X and X′are S, Y and Y′ are H, n, R₁, R₂, R₃, R₄, R₅, R_(5′), R₇, V, and T areas defined in any of the embodiments herein.

According to another embodiment in compounds of formula I, R_(5′) ishydrogen and R₅ is:

According to yet another embodiment in compounds of formula I, R_(5′) ishydrogen and R₅ is:

According to another embodiment in compounds of formula I, R_(5′) and R₅is:

According to an embodiment in compounds of formula Ia, R₅ is:

According to another embodiment in compounds of formula Ia, R₅ is:

According to another embodiment of compounds of formula I or formula Ia,the present invention provides a compound of formula IH:

wherein:

n, R₁, R₂, R₃, R₄, each R₆, R₇, V, T, X, X′, Y, and Y′ are as defined inany of the embodiments herein.

According to another embodiment for compounds of formula IH, n, R₁, R₂,R₃, R₄, R₇, V, and T are as defined in any of the embodiments herein, Xand X′ are S, Y and Y′ are H, and NR₆R₆ is:

According to another embodiment for compounds of formula IH, n, R₁, R₂,R₃, R₄, R₇, V, and T are as defined in any of the embodiments herein, Xand X′ are S, Y and Y′ are H, and NR₆R₆ is:

According to another embodiment for compounds of formula I, R₂, R₄, andR₇ are each independently H, methyl, ethyl, or propyl.

According to another embodiment for compounds of formula I, R₂, R₄, andR₇ are each H.

According to an embodiment in compounds of formula Ia, R₄ is hydrogen.

According to another embodiment in compounds of formula I or formula Ia,the present invention provides a compound of formula IJ:

wherein:

-   -   n, R₁, R₃, each R₆, V, T, X, X′, Y, and Y′ are as defined in any        of the embodiments herein.

According to another embodiment for compounds of formula IJ, n, R₁, R₃,V, and T are as defined in any of the embodiments herein, X and X′ areS, Y and Y′ are H, and NR₆R₆ is:

According to another embodiment for compounds of formula IJ, n, R₁, R₃,V, and T are as defined in any of the embodiments herein, X and X′ areS, Y and Y′ are H, and NR₆R₆ is:

According to another embodiment in compounds of formula I, R₃ is:

In another embodiment in compounds of formula I, R₃ is:

According to another embodiment in compounds of formula I, R₃ is:

According to an embodiment in compounds of formula Ia, R₃ is:

In another embodiment in compounds of formula Ia, R₃ is:

According to another embodiment in compounds of formula Ia, R₃ is:

According to another embodiment in compounds of formula I or formula Ia,the present invention provides a compound of formula IK:

wherein:

-   -   n, R₁, each R₆, V, T, X, X′, Y, and Y′ are as defined in any of        the embodiments herein.

According to another embodiment for compounds of formula IK, n, R₁, V,and T are as defined in any of the embodiments herein, X and X′ are S, Yand Y′ are H, and NR₆R₆ is:

According to another embodiment for compounds of formula IK, n, R₁, V,and T are as defined in any of the embodiments herein, X and X′ are S, Yand Y′ are H, and NR₆R₆ is:

According to another embodiment in compounds of formula I, R₁ is:

According to another embodiment in compounds of formula I, R₁ is:

In another embodiment of compounds of formula I, R₁ is cyclohexyl.

According to an embodiment in compounds of formula Ia, R₁ is:

According to another embodiment in compounds of formula Ia, R₁ is:

In another embodiment of compounds of formula Ia, R₁ is cyclohexyl.

According to another embodiment of compounds of formula I or formula Ia,the present invention provides a compound of formula IL:

wherein:

-   -   n, each R₆, V, T, X, X′, Y, and Y′ are as defined in any of the        embodiments herein.

According to another embodiment for compounds of formula IL, n, V, and Tare as defined in any of the embodiments herein, X and X′ are S, Y andY′ are H, and NR₆R₆ is:

According to another embodiment for compounds of formula IL, n, V, and Tare as defined in any of the embodiments herein, X and X′ are S, Y andY′ are H, and NR₆R₆ is:

According to another embodiment in compounds of formula I, V is O.

According to another embodiment of compounds of formula I, the presentinvention provides a compound of formula IM:

wherein:

-   -   n, each R₆, T, X, X′, Y, and Y′ are as defined in any of the        embodiments herein.

According to another embodiment for compounds of formula IM, n and T areas defined in any of the embodiments herein, X and X′ are S, Y and Y′are H, and NR₆R₆ is:

According to another embodiment for compounds of formula IM, n and T areas defined in any of the embodiments herein, X and X′ are S, Y and Y′are H, and NR₆R₆ is:

According to another embodiment in compounds of formula I, V is avalence bond.

According to another embodiment in compounds of formula I or formula Ia,the present invention provides a compound of formula IN:

wherein:

-   -   n, each R₆, T, X, X′, Y, and Y′ are as defined in any of the        embodiments herein.

According to another embodiment for compounds of formula IN, n and T areas defined in any of the embodiments herein, X and X′ are S, Y and Y′are H, and NR₆R₆ is:

According to another embodiment for compounds of formula IN, n and T areas defined in any of the embodiments herein, X and X′ are S, Y and Y′are H, and NR₆R₆ is:

According to an embodiment in compounds of formula I, T is(C3-C10)heterocyclyl- or (C5-C10)heteroaryl-;

-   -   wherein each T is optionally substituted with up to 3 J        substituents.

According to another embodiment in compounds of formula I, T is(C5-C6)heterocyclyl- or (C5-C6)heteroaryl-;

-   -   wherein each T is optionally substituted with up to 3 J        substituents.

In another embodiment in compounds of formula I, T is:

wherein:

-   -   Z is independently O, S, NR′, or C(R′)₂.

In another embodiment in compounds of formula I, T is:

In another embodiment in compounds of formula I, T is:

According to an embodiment in compounds of formula Ia, T is:

-   -   (C3-C10)heterocyclyl- or (C5-C10)heteroaryl-;        -   wherein each T is optionally substituted with up to 3 J            substituents.

According to another embodiment in compounds of formula Ia, T is(C5-C6)heterocyclyl- or (C5-C6)heteroaryl-;

-   -   wherein each T is optionally substituted with up to 3 J        substituents.

In another embodiment in compounds of formula Ia, T is:

wherein:

-   -   Z is independently O, S, NR′, or C(R′)₂.

In another embodiment in compounds of formula Ia, T is:

In another embodiment in compounds of formula Ia, T is:

According to another embodiment of compounds of formula I or formula Ia,the present invention provides a compound of formula IO:

wherein:

-   -   n, each R₆, X, X′, Y, and Y′ are as defined in any of the        embodiments herein.

According to another embodiment for compounds of formula IN, n is asdefined in any of the embodiments herein, X and X′ are S, Y and Y′ areH, and NR₆R₆ is:

According to another embodiment for compounds of formula IN, n is asdefined in any of the embodiments herein, X and X′ are S, Y and Y′ areH, and NR₆R₆ is:

According to another embodiment of compounds of formula I or formula Ia,the present invention provides a compound of formula IP:

wherein:

-   -   n, each R₆, X, X′, Y, and Y′ are as defined in any of the        embodiments herein.

According to another embodiment for compounds of formula IP, n is asdefined in any of the embodiments herein, X and X′ are S, Y and Y′ areH, and NR₆R₆ is:

According to another embodiment for compounds of formula IP, n is asdefined in any of the embodiments herein, X and X′ are S, Y and Y′ areH, and NR₆R₆ is:

According to an embodiment for compounds of formula I, said(C1-C12)-aliphatic group in R′, Y, Y′, R₁, R₂, R₃, R₄, R₆, R₇, and T is(C1-C6)-alkyl.

According to an embodiment for compounds of formula Ia, said(C1-C12)-aliphatic group in R′ and said (C1-C6)-aliphatic group in R₁,R₃, R₅, and T is (C1-C6)-alkyl.

According to another embodiment in compounds of formula I or formula Ia,the compound is:

The compounds of this invention may contain one or more asymmetriccarbon atoms and thus may occur as racemates and racemic mixtures,single enantiomers, diastereomeric mixtures and individualdiastereomers. All such isomeric forms of these compounds are expresslyincluded in the present invention. Each stereogenic carbon may be of theR or S configuration.

In one embodiment, the compounds of this invention have the structureand stereochemistry depicted in formulae ID-IP.

In another embodiment, the compounds of this invention have thestructure and stereochemistry depicted in compounds 2a to 4a.

In another embodiment, the compounds of this invention have thestructure and stereochemistry depicted in compound 7a.

Any of the embodiments recited above, including those embodiments in theabove species, may be combined to produce another embodiment of thisinvention.

As used herein, P1, P2, P3, P4 refer to the residues of an HCV proteaseinhibitor as defined in the art [J. A. Landro et al., “Mechanistic Roleof an NS4A Peptide Cofactor with the Truncated NS3 Protease of HepatitisC Virus: Elucidation of the NS4A Stimulatory Effect via Kinetic analysisand Inhibitor Mapping”, Biochemistry, 36, pp. 9340-9348 (1997)] and assuch are well known to skilled practitioners.

The present invention provides potent binders and inhibitors of the HCVNS3/NS4a serine protease. In certain embodiments of compounds offormulae I and Ia of the present invention, the compounds have P4 capsthat allow for additional hydrogen bonds with the enzyme backbone. Incertain embodiments of the present invention, a P4 cap nitrogen atom andthe carbonyl (adjacent to radical V or T in formulae I or Ia) formhydrogen bonds to the main chain carbonyl and NH groups respectively ofthe Cys-158 residue of the protease enzyme. In certain embodiments ofthe present invention, another hydrogen bond is formed by the NH moiety(represented by the N—R₂ group in formulae I and Ia, wherein R₂ ishydrogen) of the P3 group with the protease backbone. These P4 and P3hydrogen bond interactions optimize the positioning of the P4 and P3side chains in the HCV NS3/NS4a serine protease binding site.Additionally, the P2 spirocyclic proline group fills the P2 pocket andmakes favorable van der Waals contact with the Arg-155 side chain in theHCV NS3/NS4A serine protease enzyme. This invention also provides Wgroups that bind efficiently to the catalytic site of the HCV NS3/NS4aserine protease located in the P1′ pocket.

Abbreviations which are used in the schemes, preparations and theexamples that follow are:

-   THF: tetrahydrofuran-   DMF: N,N,-dimethylformamide-   EtOAc: ethyl acetate-   AcOH: acetic acid-   DMAP: dimethylaminopyridine-   HOBt: 1-hydroxybenzotriazole hydrate-   HOSu: succinic acid-   EDC: 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride-   Et₂O: diethyl ether-   BOC: tert-butyloxycarbonyl-   Cbz: benzyloxycarbonyl-   Chg: cyclohexyl glycine-   t-BG: tert-butylglycine-   DAST: (diethylamino)sulfur trifluoride-   DMSO: dimethyl sulfoxide-   DCCA: dichloroacetic acid-   DIEA: diisopropylethylamine-   MeCN: acetonitrile-   TEMPO: 2,2,6,6-tetramethyl-1-piperidinyloxy, free radical)-   DMEM: Dulbecco's Modified Eagle's Medium-   PBS: phosphate-buffered saline-   rt or RT: room temperature-   ON: overnight-   ND: not determined-   MS: mass spectrometry-   LC: liquid chromatography    General Synthetic Methodology:

The compounds of this invention may be prepared in general by methodsknown to those skilled in the art. Schemes 1A, 1B, and 1-6 belowillustrate synthetic routes to the compounds of the present invention.Other equivalent schemes, which will be readily apparent to the ordinaryskilled organic chemist, may alternatively be used to synthesize variousportions of the molecule as illustrated by the general schemes below,and the preparative examples that follow.

Scheme 1 above provides a general synthetic route for the preparation ofcompounds of formula IA wherein n, T, X, X′, R⁶, R⁵, R³, and R¹ are asdefined in any of the embodiments herein. Intermediate 11 was preparedaccording to the procedures described by Schoellkopf, et al., JustusLiebigs Ann. Chem. GE, pp. 183-202 (1976) and Stemple et al., OrganicLetters, 2(18), pp. 2769-2772 (2000). Compounds 1a-7a were preparedaccording to this scheme or variations thereof. As would be recognizedby skilled practitioners, other suitable and commercially availablecoupling reagents may be used to prepare intermediates 5, 7, 9, and 12.Additionally, it will be recognized that the commercially available Bocprotected amino acids represented by, for instance, Boc-R₃—COOH, mayalternatively be substituted with the commercial Cbz protected aminoacids. Suitable deprotection conditions to remove the Cbz protectinggroups are known to those skilled in the art. Likewise the oxidation ofintermediate 12 to compounds of formula IA can be accomplished usingother suitable conditions known to the skilled artisan.

Scheme 1A above provides a synthetic route for the preparation ofintermediate 46 from intermediate 40. Intermediate 40 was preparedaccording to the procedures described by Schoellkopf, et al., JustusLiebigs Ann. Chem. GE, pp. 183-202 (1976) and Stemple et al., OrganicLetters, 2(18), pp. 2769-2772 (2000). Esterification to the ethyl esterhydrochloride 41 was accomplished using catalytic acidic conditions. Bocprotection of the amine followed by basic hydrolysis afforded the Bocacid 43. Amine coupling with HN(R₆)₂ with EDC and succinic acid affordedamide 44 which was subsequently oxidized to the diketo amide 45 withDess-Martin periodinane. Boc removal under acidic conditions providedintermediate 46 as the hydrochloride salt wherein R₅ and R₆ are asdefined in any of the embodiments herein.

Scheme 1B above provides an alternate synthetic route for thepreparation of compounds IA from intermediate 46.

Scheme 2 above provides a general synthetic route for the preparation ofcompounds of formula IB wherein T, R⁶, R⁵, R³, and R¹ are as defined inany of the embodiments herein. As would be recognized by skilledpractitioners, other suitable and commercially available couplingreagents may be used to prepare intermediates 15, 17, 19, and 21.Additionally, it will be recognized that the commercially available Bocprotected amino acids represented by, for instance, Boc-R₃—COOH, mayalternatively be substituted with the commercial Cbz protected aminoacids. Suitable deprotection conditions to remove the Cbz protectinggroups are known to those skilled in the art. Likewise the oxidation ofintermediate 21 to compounds of formula IB may be accomplished usingother suitable conditions known to the skilled artisan. One of skill inthe art will also recognize that compounds of formula IB may also beprepared from intermediate 46 using the conditions described above inScheme 1B.

Scheme 3 above provides a general synthetic route for the preparation ofcompounds of formula IC wherein X, X′, T, R⁶, R⁵, R³, and R¹ are asdefined in any of the embodiments herein. One of skill in the art willalso recognize that compounds of formula IC may also be prepared fromintermediate 46 using the conditions described above in Scheme 1B.

Scheme 4 above provides a synthetic route for the preparation of pyrroleacid intermediate 5b. It will be appreciated by those skilled in the artthat other pyrrole analogs of interest may be synthesized bymodifications of scheme 4.

Scheme 5 above provides a synthetic route for the preparation ofcompound 2a. Compounds 3a, 5a, 6a, and 7a were also prepared generallyaccording to scheme 5. One of skill in the art will also recognize thatcompounds of formula 2a, 3a, 5a, 6a, and 7a may also be prepared fromintermediate 46 using the conditions described above in Scheme 1B.

Scheme 6 above provides a synthetic route for the preparation ofcompound 1a. Compound 4a was also prepared generally according to scheme6. One of skill in the art will also recognize that compounds of formula1a may also be prepared from intermediate 46 using the conditionsdescribed above in Scheme 1B.

Although certain exemplary embodiments are depicted and described below,it will be appreciated that compounds of this invention can be preparedaccording to the methods described generally above using appropriatestarting materials generally available to one of ordinary skill in theart.

Another embodiment of this invention provides a pharmaceuticalcomposition comprising a compound of formula I or formula Ia or apharmaceutically acceptable salt thereof. According to one embodiment,the compound of formula I or formula Ia is present in an amounteffective to decrease the viral load in a sample or in a patient,wherein said virus encodes a serine protease necessary for the virallife cycle, and a pharmaceutically acceptable carrier.

If pharmaceutically acceptable salts of the compounds of this inventionare utilized in these compositions, those salts are preferably derivedfrom inorganic or organic acids and bases. Included among such acidsalts are the following: acetate, adipate, alginate, aspartate,benzoate, benzene sulfonate, bisulfate, butyrate, citrate, camphorate,camphor sulfonate, cyclopentane-propionate, digluconate, dodecylsulfate,ethanesulfonate, fumarate, glucoheptanoate, glycerophosphate,hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide,hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate,methanesulfonate, 2-naphthalenesulfonate, nicotinate, oxalate, pamoate,pectinate, persulfate, 3-phenyl-propionate, picrate, pivalate,propionate, succinate, tartrate, thiocyanate, tosylate and undecanoate.Base salts include ammonium salts, alkali metal salts, such as sodiumand potassium salts, alkaline earth metal salts, such as calcium andmagnesium salts, salts with organic bases, such as dicyclohexylaminesalts, N-methyl-D-glucamine, and salts with amino acids such asarginine, lysine, and so forth.

Also, the basic nitrogen-containing groups may be quaternized with suchagents as lower alkyl halides, such as methyl, ethyl, propyl, and butylchloride, bromides and iodides; dialkyl sulfates, such as dimethyl,diethyl, dibutyl and diamyl sulfates, long chain halides such as decyl,lauryl, myristyl and stearyl chlorides, bromides and iodides, aralkylhalides, such as benzyl and phenethyl bromides and others. Water oroil-soluble or dispersible products are thereby obtained.

The compounds utilized in the compositions and methods of this inventionmay also be modified by appending appropriate functionalities to enhanceselective biological properties. Such modifications are known in the artand include those which increase biological penetration into a givenbiological system (e.g., blood, lymphatic system, central nervoussystem), increase oral availability, increase solubility to allowadministration by injection, alter metabolism and alter rate ofexcretion.

Pharmaceutically acceptable carriers that may be used in thesecompositions include, but are not limited to, ion exchangers, alumina,aluminum stearate, lecithin, serum proteins, such as human serumalbumin, buffer substances such as phosphates, glycine, sorbic acid,potassium sorbate, partial glyceride mixtures of saturated vegetablefatty acids, water, salts or electrolytes, such as protamine sulfate,disodium hydrogen phosphate, potassium hydrogen phosphate, sodiumchloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sodiumcarboxymethylcellulose, polyacrylates, waxes,polyethylene-polyoxypropylene-block polymers, polyethylene glycol andwool fat.

According to one embodiment, the compositions of this invention areformulated for pharmaceutical administration to a mammal, preferably ahuman being.

Such pharmaceutical compositions of the present invention may beadministered orally, parenterally, by inhalation spray, topically,rectally, nasally, buccally, vaginally or via an implanted reservoir.The term “parenteral” as used herein includes subcutaneous, intravenous,intramuscular, intra-articular, intra-synovial, intrasternal,intrathecal, intrahepatic, intralesional and intracranial injection orinfusion techniques. Preferably, the compositions are administeredorally or intravenously.

Sterile injectable forms of the compositions of this invention may beaqueous or oleaginous suspension. These suspensions may be formulatedaccording to techniques known in the art using suitable dispersing orwetting agents and suspending agents. The sterile injectable preparationmay also be a sterile injectable solution or suspension in a non-toxicparenterally acceptable diluent or solvent, for example as a solution in1,3-butanediol. Among the acceptable vehicles and solvents that may beemployed are water, Ringer's solution and isotonic sodium chloridesolution. In addition, sterile, fixed oils are conventionally employedas a solvent or suspending medium. For this purpose, any bland fixed oilmay be employed including synthetic mono- or di-glycerides. Fatty acids,such as oleic acid and its glyceride derivatives are useful in thepreparation of injectables, as are natural pharmaceutically-acceptableoils, such as olive oil or castor oil, especially in theirpolyoxyethylated versions. These oil solutions or suspensions may alsocontain a long-chain alcohol diluent or dispersant, such ascarboxymethyl cellulose or similar dispersing agents which are commonlyused in the formulation of pharmaceutically acceptable dosage formsincluding emulsions and suspensions. Other commonly used surfactants,such as Tweens, Spans and other emulsifying agents or bioavailabilityenhancers which are commonly used in the manufacture of pharmaceuticallyacceptable solid, liquid, or other dosage forms may also be used for thepurposes of formulation.

Dosage levels of between about 0.01 and about 100 mg/kg body weight perday, preferably between about 0.5 and about 75 mg/kg body weight per dayof the protease inhibitor compounds described herein are useful in amonotherapy for the prevention and treatment of antiviral, particularlyanti-HCV mediated disease. Typically, the pharmaceutical compositions ofthis invention will be administered from about 1 to about 5 times perday or alternatively, as a continuous infusion. Such administration canbe used as a chronic or acute therapy. The amount of active ingredientthat may be combined with the carrier materials to produce a singledosage form will vary depending upon the host treated and the particularmode of administration. A typical preparation will contain from about 5%to about 95% active compound (w/w). Preferably, such preparationscontain from about 20% to about 80% active compound.

When the compositions of this invention comprise a combination of acompound of formula I or formula Ia and one or more additionaltherapeutic or prophylactic agents, both the compound and the additionalagent should be present at dosage levels of between about 10 to 100%,and more preferably between about 10 to 80% of the dosage normallyadministered in a monotherapy regimen.

The pharmaceutical compositions of this invention may be orallyadministered in any orally acceptable dosage form including, but notlimited to, capsules, tablets, aqueous suspensions or solutions. In thecase of tablets for oral use, carriers that are commonly used includelactose and corn starch. Lubricating agents, such as magnesium stearate,are also typically added. For oral administration in a capsule form,useful diluents include lactose and dried cornstarch. When aqueoussuspensions are required for oral use, the active ingredient is combinedwith emulsifying and suspending agents. If desired, certain sweetening,flavoring or coloring agents may also be added.

Alternatively, the pharmaceutical compositions of this invention may beadministered in the form of suppositories for rectal administration.These may be prepared by mixing the agent with a suitable non-irritatingexcipient that is solid at room temperature but liquid at rectaltemperature and therefore will melt in the rectum to release the drug.Such materials include cocoa butter, beeswax and polyethylene glycols.

The pharmaceutical compositions of this invention may also beadministered topically, especially when the target of treatment includesareas or organs readily accessible by topical application, includingdiseases of the eye, the skin, or the lower intestinal tract. Suitabletopical formulations are readily prepared for each of these areas ororgans.

Topical application for the lower intestinal tract may be effected in arectal suppository formulation (see above) or in a suitable enemaformulation. Topically-transdermal patches may also be used.

For topical applications, the pharmaceutical compositions may beformulated in a suitable ointment containing the active componentsuspended or dissolved in one or more carriers. Carriers for topicaladministration of the compounds of this invention include, but are notlimited to, mineral oil, liquid petrolatum, white petrolatum, propyleneglycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax andwater. Alternatively, the pharmaceutical compositions may be formulatedin a suitable lotion or cream containing the active components suspendedor dissolved in one or more pharmaceutically acceptable carriers.Suitable carriers include, but are not limited to, mineral oil, sorbitanmonostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol,2-octyldodecanol, benzyl alcohol and water.

For ophthalmic use, the pharmaceutical compositions may be formulated asmicronized suspensions in isotonic, pH adjusted sterile saline, or,preferably, as solutions in isotonic, pH adjusted sterile saline, eitherwith our without a preservative such as benzylalkonium chloride.Alternatively, for ophthalmic uses, the pharmaceutical compositions maybe formulated in an ointment such as petrolatum.

The pharmaceutical compositions of this invention may also beadministered by nasal aerosol or inhalation. Such compositions areprepared according to techniques well known in the art of pharmaceuticalformulation and may be prepared as solutions in saline, employing benzylalcohol or other suitable preservatives, absorption promoters to enhancebioavailability, fluorocarbons, and/or other conventional solubilizingor dispersing agents.

In another embodiment, the pharmaceutical compositions are formulatedfor oral administration.

In one embodiment, the compositions of this invention additionallycomprise another agent, such as a cytochrome P-450 inhibitor. Suchcytochrome P-450 inhibitors include, but are not limited to, ritonavir.

If an embodiment of this invention involves a CYP inhibitor, any CYPinhibitor that improves the pharmacokinetics of the relevant NS3/4Aprotease may be used in a method of this invention. These CYP inhibitorsinclude, but are not limited to, ritonavir (WO 94/14436), ketoconazole,troleandomycin, 4-methyl pyrazole, cyclosporin, clomethiazole,cimetidine, itraconazole, fluconazole, miconazole, fluvoxamine,fluoxetine, nefazodone, sertraline, indinavir, nelfinavir, amprenavir,fosamprenavir, saquinavir, lopinavir, delavirdine, erythromycin, VX-944,and VX-497. According to one embodiment, the CYP inhibitors includeritonavir, ketoconazole, troleandomycin, 4-methyl pyrazole, cyclosporin,and clomethiazole.

Methods for measuring the ability of a compound to inhibit cytochromeP50 monooxygenase activity are known (see U.S. Pat. No. 6,037,157 andYun, et al. Drug Metabolism & Disposition, vol. 21, pp. 403-407 (1993).

A CYP inhibitor employed in this invention may be an inhibitor of onlyone isozyme or more than one isozyme. If the CYP inhibitor inhibits moreisozyme, the inhibitor may nevertheless inhibit one isozyme moreselectively than another isozyme. Any such CYP inhibitors may be used ina method of this invention.

In a method of this invention, the CYP inhibitor may be administeredtogether with the Hepatitis C virus NS3/4A protease inhibitor in thesame dosage form or in separate dosage forms.

If the CYP inhibitor and protease inhibitor are administered in separatedosage forms, each inhibitor may be administered about simultaneously.Alternatively, the CYP inhibitor may be administered in any time periodaround administration of the protease inhibitor. That is, the CYPinhibitor may be administered prior to, together with, or following theNS3/4A protease inhibitor. The time period of administration should besuch that the CYP inhibitor affects the metabolism of the proteaseinhibitor. For example, if the protease inhibitor is administered first,the CYP inhibitor should be administered before the protease inhibitoris substantially metabolized and/or excreted (e.g., within the half-lifeof the protease inhibitor).

In another embodiment, the compositions of this invention additionallycomprise another anti-viral agent, preferably an anti-HCV agent. Suchanti-viral agents include, but are not limited to, immunomodulatoryagents, such as α-, β-, and γ-interferons, pegylated derivatizedinterferon-α compounds, and thymosin; other anti-viral agents, such asribavirin, amantadine, and telbivudine; other inhibitors of hepatitis Cproteases (NS2-NS3 inhibitors and NS3-NS4A inhibitors); inhibitors ofother targets in the HCV life cycle, including metalloprotease, helicaseand polymerase inhibitors; inhibitors of internal ribosome entry;broad-spectrum viral inhibitors, such as IMPDH inhibitors (e.g.,compounds of U.S. Pat. Nos. 5,807,876, 6,498,178, 6,344,465, 6,054,472,WO 97/40028, WO 98/40381, WO 00/56331, and mycophenolic acid andderivatives thereof, and including, but not limited to VX-497, VX-148,and/or VX-944); or combinations of any of the above.

The term “interferon” as used herein means a member of a family ofhighly homologous species-specific proteins that inhibit viralreplication and cellular proliferation, and modulate immune response,such as interferon alpha, interferon beta, or interferon gamma. TheMerck Index, entry 5015, Twelfth Edition.

Upon improvement of a patient's condition, a maintenance dose of acompound, composition or combination of this invention may beadministered, if necessary. Subsequently, the dosage or frequency ofadministration, or both, may be reduced, as a function of the symptoms,to a level at which the improved condition is retained when the symptomshave been alleviated to the desired level, treatment should cease.Patients may, however, require intermittent treatment on a long-termbasis upon any recurrence of disease symptoms.

It should also be understood that a specific dosage and treatmentregimen for any particular patient will depend upon a variety offactors, including the activity of the specific compound employed, theage, body weight, general health, sex, diet, time of administration,rate of excretion, drug combination, and the judgment of the treatingphysician and the severity of the particular disease being treated. Theamount of active ingredients will also depend upon the particulardescribed compound and the presence or absence and the nature of theadditional anti-viral agent in the composition.

According to another embodiment, the invention provides a method fortreating a patient infected with a virus characterized by a virallyencoded serine protease that is necessary for the life cycle of thevirus by administering to said patient a pharmaceutically acceptablecomposition of this invention. Preferably, the methods of this inventionare used to treat a patient suffering from a HCV infection. Suchtreatment may completely eradicate the viral infection or reduce theseverity thereof. More preferably, the patient is a human being.

In an alternate embodiment, the methods of this invention additionallycomprise the step of administering to said patient an anti-viral agentpreferably an anti-HCV agent. Such anti-viral agents include, but arenot limited to, immunomodulatory agents, such as α-, β-, andγ-interferons, pegylated derivatized interferon-α compounds, andthymosin; other anti-viral agents, such as ribavirin, amantadine, andtelbivudine; other inhibitors of hepatitis C proteases (NS2-NS3inhibitors and NS3-NS4A inhibitors); inhibitors of other targets in theHCV life cycle, including metalloprotease, helicase and polymeraseinhibitors; inhibitors of internal ribosome entry; broad-spectrum viralinhibitors, such as IMPDH inhibitors (e.g., compounds of U.S. Pat. Nos.5,807,876, 6,498,178, 6,344,465, 6,054,472, WO 97/40028, WO 98/40381, WO00/56331, and mycophenolic acid and derivatives thereof, and including,but not limited to VX-497, VX-148, and/or VX-944); or combinations ofany of the above.

Such additional agent may be administered to said patient as part of asingle dosage form comprising both a compound of this invention and anadditional anti-viral agent. Alternatively the additional agent may beadministered separately from the compound of this invention, as part ofa multiple dosage form, wherein said additional agent is administeredprior to, together with or following a composition comprising a compoundof this invention.

In yet another embodiment the present invention provides a method ofpre-treating a biological substance intended for administration to apatient comprising the step of contacting said biological substance witha pharmaceutically acceptable composition comprising a compound of thisinvention. Such biological substances include, but are not limited to,blood and components thereof such as plasma, platelets, subpopulationsof blood cells and the like; organs such as kidney, liver, heart, lung,etc; sperm and ova; bone marrow and components thereof, and other fluidsto be infused into a patient such as saline, dextrose, etc.

According to another embodiment the invention provides methods oftreating materials that may potentially come into contact with a viruscharacterized by a virally encoded serine protease necessary for itslife cycle. This method comprises the step of contacting said materialwith a compound according to the invention. Such materials include, butare not limited to, surgical instruments and garments (e.g. clothes,gloves, aprons, gowns, masks, eyeglasses, footwear, etc.); laboratoryinstruments and garments (e.g. clothes, gloves, aprons, gowns, masks,eyeglasses, footwear, etc.); blood collection apparatuses and materials;and invasive devices, such as shunts, stents, etc.

In another embodiment, the compounds of this invention may be used aslaboratory tools to aid in the isolation of a virally encoded serineprotease. This method comprises the steps of providing a compound ofthis invention attached to a solid support; contacting said solidsupport with a sample containing a viral serine protease underconditions that cause said protease to bind to said solid support; andeluting said serine protease from said solid support. Preferably, theviral serine protease isolated by this method is HCV NS3-NS4A protease.

In order that this invention be more fully understood, the followingpreparative and testing examples are set forth. These examples are forthe purpose of illustration only and are not to be construed as limitingthe scope of the invention in any way.

EXAMPLES

¹H-NMR spectra were recorded at 500 MHz using a Bruker AMX 500instrument. Mass spec. samples were analyzed on a MicroMass ZQ orQuattro II mass spectrometer operated in single MS mode withelectrospray ionization. Samples were introduced into the massspectrometer using flow injection (FIA) or chromatography. Mobile phasefor all mass spec. analysis consisted of acetonitrile-water mixtureswith 0.2% formic acid as a modifier.

As used herein, the term “R_(t)(min)” refers to the HPLC retention time,in minutes, associated with the compound. The HPLC retention timeslisted were either obtained from the mass spec. data or using thefollowing method:

-   Instrument: Hewlett Packard HP-1050;-   Column: YMC C₁₈ (Cat. No. 326289C46);-   Gradient/Gradient Time: 10-90% CH₃CN/H2O over 9 minutes, then 100%    CH₃CN for 2 minutes;-   Flow Rate: 0.8 ml/min;-   Detector Wavelength: 215 nM and 245 nM.

Chemical naming for selected compounds herein was accomplished using thenaming program provided by CambridgeSoft Corporations ChemDraw Ultra®,version 7.0.1.

Example 1 3-acetyl-4,5-dimethyl-2-pyrrole carboxylic acid (5b)

A solution of sodium nitrite (36.9 g, 0.534 mol) in 70 mL of water wasadded dropwise to a stirred solution of ethylacetoacetate (70 g, 0.538mol) in 1401 mL of glacial acetic acid at 0° C. After the addition wascomplete, the light yellow reaction mixture was allowed to warm to roomtemperature. After 30 minutes, all the starting material had beenconsumed, the reaction was quenched with 350 mL of water and extractedwith ethyl acetate (2×125 mL). The organic extracts were combined andwashed with water (2×125 mL) and saturated sodium hydrogen carbonateaqueous solution (2×105 mL). The organic layer was dried with sodiumsulfate and concentrated in vacuo to give 84.2 g (98%) ofethyl-2-Hydroxyimino-3-oxobutanoate 1b as a pale yellow oil. ¹H NMR(CDCl₃) δ 10.3 (s, 1H), 4.2 (q, 2H), 2.3 (s, 3H), 1.3 (t, 3H) ppm.

Crushed sodium (12.4 g, 0.540 mol) was added to a solution of 2-butanone(48.2 mL, 0.538 mol) and ethyl formate (43.47 mL, 0.538 mol) in dryether (540 mL) with vigorous mechanical stirring over a period of 1 h,during which time the mixture was chilled in an ice-salt bath. Themixture was then stirred at room temp. for 14 hours. After cooling thereaction mixture to 4° C. for a few hours, the precipitated sodium saltwas obtained by filtration and washed thoroughly with cold, dry ether toafford 49.3 g (75%) of the desired sodium salt of2-methy-3-oxobutyraldehyde 2b. ¹H NMR (DMSO-d₆) δ 9.1 (s, 1H), 1.9 (s,3H), 1.3 (s, 3H) ppm.

Sodium salt 2b (49.3 g, 0.404 mol) and oxime 1b (64.23, 0.404 mol) werestirred in 300 mL of 70% acetic acid/30% water and warmed to 50° C. Zincpowder (42.21 g, 0.646 mol) was added portion-wise over 30 minutesmaintaining the temperature below 100° C. When the addition wascomplete, the suspension was refluxed for 15 minutes, then poured into 4L of ice-water. After a short time, the product precipitated out togive, after filtration, 30.1 g (45%) of the desiredethyl-4,5-dimethyl-2-pyrrole carboxylate 3b. ¹H NMR (CDCl₃) δ 9.0 (bs,1H), 6.7 (s, 1H), 4.3 (q, 2H), 2.3 (s, 3H), 2.0 (s, 3H), 1.3 (t, 3H)ppm.

To a solution of aluminum chloride (50.19 g, 0.376 mol) in drydichloroethane (580 mL) at 25° C. was added slowly acetic anhydride(17.75 mL, 0.188 mol). The resulting mixture was stirred at room temp.for 10 minutes, then a solution of pyrrole 3b (10.49 g, 0.0627 mol) indichloroethane (30 mL) was added and the reaction mixture was stirred atroom temp. for 2 hours. After an additional 3 hours at 80° C., themixture was poured into ice water and extracted with dichloromethane.The organic layer was dried with anhydrous sodium sulfate andconcentrated in vacuo to an orange residue. Short plug filtration oversilica gel (30% ethyl acetate/70% hexanes) gave 7.5 g (60%) ofethyl-3-acetyl-4,5-dimethyl-2-pyrrole carboxylate 4b. ¹H NMR (CDCl₃) δ9.0 (bs, 1H), 4.3 (q, 2H), 2.7 (s, 3H), 2.1 (s, 3H), 1.9 (s, 3H), 1.3(t, 3H) ppm.

A mixture of pyrrole ester 4b (8.2 g, 0.0392 mol), in ethanol and 100 mLof 10% potassium hydroxide were refluxed for 1 hour. The mixture wascooled and concentrated in vacuo to an oil. Water was added to the oil,the mixture acidified with dilute HCl and extracted with ether. Theorganic phase was dried with anh. sodium sulfate and concentrated invacuo to a solid residue. The compound was recrystallized in 80 mL ofethanol to give 5.8 g of pure 3-acetyl-4,5-dimethyl-2-pyrrole carboxylicacid 5b as a solid. ¹H NMR (DMSO-d₆) δ 2.5 (s, 3H), 2.2 (s, 3H), 2.0 (s,3H) ppm.

Example 22-(2-{2-cyclohexyl-2-[(pyrazine-2-carbonyl)-amino]-acetylamino}-3,3-dimethyl-butyryl)-6,10-dithia-2-azaspiro[4.5]decane-3-carboxylicacid(1-cyclopropylaminooxalyl-butyl)-amide (2a)

To a solution of L-4-hydroxy-pyrrolidine-1,2-dicarboxylic acid 1-benzylester 2-methyl ester 1 (3.0 g, 1.0 eq, Advanced Chem Tech) in toluene(30 mL)/ethyl acetate (30 mL) was added NaBr (1.28 g, 1.14 eq) in water(5 mL). TEMPO (17 mg) was added, the mixture cooled to 4° C. and asolution of Clorox® (18 mL), sodium bicarbonate (2.75 g) and water (to40 mL total volume) was added over 30 minutes. The resulting suspensionwas stirred 10 minutes before adding isopropanol (0.2 mL). The organicphase was separated and the aqueous phase extracted with ethyl acetate.The combined organic layers were washed with a 0.3 N solution of sodiumthiosulfate and then brine, dried over sodium sulfate, filtered, andconcentrated in vacuo to an amber oil. Purification via a silica gelplug eluting with an EtOAc/hexanes gradient (100% hexanes to 40%EtOAc/hexanes in 10% steps) afforded 2.85 g (96%) of4-oxo-pyrrolidine-1,2-dicarboxylic acid 1-benzyl ester 2-methyl ester 2as a clear oil. ¹H NMR (CDCl₃) δ 7.35 (m, 5H), 5.15-5.30 (m, 2H),4.80-4.90 (m, 1H), 3.95-4.05 (m, 2H), 3.80 (s, 3/5 of 3H), 3.65 (s, 2/5of 3H), 3.0 (m, 1H), 2.65 (d, 3/5 of 1H), 2.60 (d, 2/5 of 1H) ppm.

Ketone 2 (250 mg) in CH₂Cl₂ at 15° C. was treated with1,3-propanedithiol (100 uL) dropwise followed by BF₃OEt₂ (119 uL). Themixture was warmed to room temperature and stirred overnight. Thereaction was quenched by adding 1 mL potassium carbonate aqueoussolution (2 g/30 mL) followed by 321 uL of saturated sodium bicarbonateto adjust to pH 7-8. Washed organics with water, brine, then dried overanhydrous magnesium sulfate, filtered and concentrated in vacuo.Purification on a plug of silica gel eluting with toluene→hexane/ethylether (2:3→0:1) yielded 200 mg (60%) of desired6,10-dithia-2-aza-spiro[4.5]decane-2,3-dicarboxylic acid-2-benzyl ester3-methyl ester 26 as a clear oil. ¹H NMR (CDCl₃) δ 7.30 (m, 5H),5.05-5.25 (m, 2H), 4.6 (t, 0.5H), 4.55 (t, 0.5H), 3.8 (s, 1.5H), 3.75(m, 1H), 3.6 (s, 1.5H), 2.95 (m, 1H), 2.85 (m, 3H), 2.75 (m, 1H), 2.4(m, 0.5H), 2.35 (m, 0.5H), 2.0 (m, 2H) ppm.

Cbz-protected dithiane (50 mg) 26 in AcOH (140 uL) was treated with 30%HBr/AcOH (210 uL) and stirred for 2 hours at room temperature. Ethylether (20 mL) was added, the suspension stirred, solvent decanted offand then the procedure repeated twice more to give 40 mg (95%) ofdesired 6,10-dithia-2-aza-spiro[4.5]decane-3-carboxylic acid methylester 27 as a reddish-brown solid and as the HBr salt. ¹H NMR (CDCl₃) δ4.75 (t, 1H), 3.8 (s, 3H), 3.65 (d, 2H), 2.9-3.1 (m, 4H), 2.7 (m,1H),2.55 (m, 1H), 1.95 (m, 1H), 1.85 (m, 1H) ppm.

L-Boc-tert-butyl glycine (243 mg, Bachem), EDC (201 mg), HOBt (161 mg),and DIEA (502 uL) in DMF (3 mL) was treated with the amine salt 27 (300mg) in DMF (1 mL) and stirred at room temp. overnight. The mixture waspartitioned between ethyl acetate and 1.0 N HCl, the organics washedwith saturated sodium bicarbonate, 1.0 N glycine sodium salt solution,10% potassium carbonate solution, and brine then dried over anhydroussodium sulfate, filtered, and concentrated in vacuo. Purification on aplug of silica gel eluting with 30% EtOAc/hexanes afforded 300 mg (70%)of desired2-(2-tert-butoxycarbonylamino-3,3-dimethyl-butyryl)-6,10-dithia-2-aza-spiro[4.5]decane-3-carboxylicacid methyl ester 28 as a white solid. ¹H NMR (CDCl₃) δ 5.2 (d, 1H), 4.7(t, 1H), 4.65 (s, 1H), 4.3 (d, 1H), 3.8 (d, 1H), 3.75 (s, 3H), 3.1 (m,1H), 3.0 (m, 1H), 2.8 (m, 1H), 2.75 (m, 1H), 2.6 (m, 1H), 2.2 (m, 1H),2.1 (m, 1H), 1.95 (m, 1H), 1.45 (s, 9H), 1.05 (s, 9H) ppm.

Boc protected amine 28 (243 mg) in dioxane (1 mL) was treated with a4.ON HCl/dioxane solution (2 mL) and stirred for 2 hours at roomtemperature. The mixture was concentrated in vacuo, slurried in CH₂Cl₂and evaporated in vacuo to give 208 mg (100%) of desired2-(2-amino-3,3-dimethyl-butyryl)-6,10-dithia-2-aza-spiro[4.5]decane-3-carboxylicacid methyl ester 29 as a white solid and as the HCl salt. Mass Spec.MH+=347.1.

L-Boc-cyclohexyl glycine (154 mg, Bachem), EDC (115 mg), HOBt (81 mg),and DIEA (284 mg) in CH₂Cl₂ (1 mL) was treated with amine salt 29 (189mg) in CH₂Cl₂ (2 mL) and the mixture stirred for 2 hours. The mixturewas partitioned between ethyl acetate and 1.0 N HCl, the organics washedwith sodium bicarbonate, 1.0 N glycine sodium salt solution, 10%potassium carbonate solution and brine then dried over sodium sulfate,filtered, and concentrated in vacuo. Purification on a plug of silicagel eluting with 30% EtOAc/hexanes afforded 221 mg (70%) of2-[2-tert-butoxycarbonylamino-2-cyclohexyl-acetylamino)-3,3-dimethyl-butyryl]-6,10-dithia-2-aza-spiro[4.5]decane-3-carboxylicacid methyl ester 30 as a white solid.

¹H NMR (CDCl₃) δ 6.4 (d, 1H), 5.0 (d, 1H), 4.7 (m, 2H), 4.6 (d, 1H), 3.9(m, 1H), 3.8 (d, 1H), 3.7 s, 1H), 3.0 (m, 2H), 2.8 (m, 1H), 2.7 (m, 1H),2.6 (m, 1H), 2.2 (m, 1H), 2.15 (m, 1H), 2.0 (m, 1H), 1.65 (m, 7H), 1.45(s, 9H), 1.15 (m, 4H), 1.05 (s, 9H) ppm.

Boc protected amine 30 (221 mg) in dioxane (1 mL) was treated with a4.0N HCl/dioxane solution (2 mL) and stirred for 2 hours at roomtemperature. The mixture was concentrated in vacuo, slurried in CH₂Cl₂,evaporated, the procedure repeated and the mixture evaporated in vacuoto give 197 mg (100%) of desired2-[2-(2-amino-2-cyclohexyl-acetylamino)-3,3-dimethyl-butyryl]-6,10-dithia-2-aza-spiro[4.5]decane-3-carboxylicacid methyl ester 31 as a white solid and as the HCl salt. Mass spec.MH+=486.2.

Pyrazine acid 32 (26 mg, Aldrich Chem Co.), EDC (40 mg), HOBt (32 mg),and DIEA (99 uL) in CH₂Cl₂ (2 mL) was treated with amine salt (98 mg) inCH₂Cl₂ (2 mL) and stirred at RT for 3 hours. The mixture was partitionedbetween EtOAc and 1.0N HCl washed with brine then dried over sodiumsulfate, filtered, and concentrated in vacuo. Purification on a plug ofsilica gel eluting with 100% EtOAc afforded 50 mg (45%) of desired2-[2-{2-cyclohexyl-2-[(pyrazine-2-carbonyl)-amino]-acetylamino}-3,3-dimethyl-butyryl)-6,10-dithia-2-aza-spiro[4.5]decane-3-carboxylicacid methyl ester 33 as a white solid. Mass spec. MH+=592.1, MH−=590.2.

Ester 33 (50 mg) in THF-water (400 uL-100 uL) was treated with LiOH (7mg) and the mixture stirred for 3 hours at room temperature. The mixturewas evaporated, diluted with EtOAc, washed with 1.0N HCl and brine thendried over magnesium sulfate, filtered, and concentrated in vacuo togive2-[2-{2-cyclohexyl-2-[(pyrazine-2-carbonyl)-amino]-acetylamino}-3,3-dimethyl-butyryl)-6,10-dithia-2-aza-spiro[4.5]decane-3-carboxylicacid 34 as a white solid which was used in the next step without furtherpurification. Mass Spec. MH+=578.0, MH−=576.2.

Acid 34 (49 mg), EDC (14.2 mg), HOBt (17.8 mg), and DIEA (44 uL) inCH₂Cl₂ (1 mL) was treated with 3-amino-2-hydroxy-hexanoic acidcyclopropylamine 35 (17.3 mg, prepared according to the methodsdescribed by U. Schoellkopf et al., Justus Liebigs Ann. Chem. GE, 1976,183-202, and J. Stemple et al., Organic Letters 2000, 2(18), 2769-2772)in CH₂Cl₂ (1 mL) and the mixture stirred overnight at RT. EtOAc wasadded, the organics washed with 1.0N HCl and brine, then dried oversodium sulfate, filtered, and concentrated in vacuo. Purification on aplug of silica gel eluting with 2% MeOH/CH₂Cl₂ afforded 31 mg (50%) ofdesired2-[2-{2-cyclohexyl-2-[(pyrazine-2-carbonyl)-amino]-acetylamino}-3,3-dimethyl-butyryl)-6,10-dithia-2-aza-spiro[4.5]decane-3-carboxylicacid[1-(cyclopropylaminooxalyl-butyl)-amide 36 as a white solid. Massspec. MH+=746.1, MH−=744.3.

Hydroxyamide 36 (31 mg) in EtOAc (620 uL) was treated with EDC (120 mg)followed by DMSO (233 uL), then dichloroacetic acid (34 uL) and themixture stirred for 30 minutes at room temp. The reaction mixture wasdiluted with 1.0N HCl (620 uL), the organics washed with water, thenconcentrated in vacuo and purified by preparative HPLC to give 14 mg(45%) of desired2-(2-{2-cyclohexyl-2-[(pyrazine-2-carbonyl)-amino]-acetylamino}-3,3-dimethyl-butyryl)-6,10-dithia-2-aza-spiro[4.5]decane-3-carboxylicacid(1-cyclopropylaminooxalyl-butyl)-amide 2a as a white solid. ¹H NMR(CDCl₃) δ 9.4 (s, 1H), 8.75 (s, 1H), 8.65 (s, 1H), 8.3 (s, 1H), 7.45 (d,1H), 6.8 (d, 1H), 5.4 (d, 1H), 4.8 (m, 2H), 4.6 (m, 1H), 4.5 (m, 1H),3.7 (d, 1H), 3.1 (m, 2H), 2.8 (m, 2H), 2.65 (m, 2H), 2.3 (m, 1H), 2.2(m, 1H), 1.95 (m, 3H), 1.7 (m, 6H), 1.4 (m, 2H), 1.15 (m, 4H), 1.05 (s,9H), 0.9 (m, 4H), 0.85 (m, 2H), 0.65 (m, 2H) ppm.

Example 32-(2-{2-[(3-acetyl-4,5-dimethyl-1H-pyrrole-2-carbonyl)-amino]-2-cyclohexyl-acetylamino}-3,3-dimethyl-butyryl)-6,10-dithia-2-aza-spiro[4.5]decane-3-carboxylicacid(1-cyclopropylaminooxalyl-butyl)-amide (1a)

This compound was prepared from2-[2-(2-amino-2-cyclohexyl-acetylamino)-3,3-dimethyl-butyryl]-6,10-dithia-2-aza-spiro[4.5]decane-3-carboxylicacid methyl ester 31 (prepared as described above in example 2) and3-acetyl-4,5-dimethyl-2-pyrrole carboxylic acid 5b (prepared asdescribed above in example 1) using procedures similar to thosedescribed in example 2. The title compound was isolated as a white solid(11% for last step). LCMS: retention time=4.8 min, M+H=801.2.

Example 4 HCV Replicon Cell Assay Protocol

Cells containing hepatitis C virus (HCV) replicon were maintained inDMEM containing 10% fetal bovine serum (FBS), 0.25 mg per ml of G418,with appropriate supplements (media A).

On day 1, replicon cell monolayer was treated with a trypsin:EDTAmixture, removed, and then media A was diluted into a finalconcentration of 100,000 cells per ml wit. 10,000 cells in 100 ul wereplated into each well of a 96-well tissue culture plate, and culturedovernight in a tissue culture incubator at 37° C.

On day 2, compounds (in 100% DMSO) were serially diluted into DMEMcontaining 2% FBS, 0.5% DMSO, with appropriate supplements (media B).The final concentration of DMSO was maintained at 0.5% throughout thedilution series.

Media on the replicon cell monolayer was removed, and then media Bcontaining various concentrations of compounds was added. Media Bwithout any compound was added to other wells as no compound controls.

Cells were incubated with compound or 0.5% DMSO in media B for 48 hoursin a tissue culture incubator at 37° C. At the end of the 48-hourincubation, the media was removed, and the replicon cell monolayer waswashed once with PBS and stored at −80° C. prior to RNA extraction.

Culture plates with treated replicon cell monolayers were thawed, and afixed amount of another RNA virus, such as Bovine Viral Diarrhea Virus(BVDV) was added to cells in each well. RNA extraction reagents (such asreagents from RNeasy kits) were added to the cells immediately to avoiddegradation of RNA. Total RNA was extracted according the instruction ofmanufacturer with modification to improve extraction efficiency andconsistency. Finally, total cellular RNA, including HCV replicon RNA,was eluted and stored at −80° C. until further processing.

A Taqman real-time RT-PCR quantification assay was set up with two setsof specific primers and probe. One was for HCV and the other was forBVDV. Total RNA extractants from treated HCV replicon cells was added tothe PCR reactions for quantification of both HCV and BVDV RNA in thesame PCR well. Experimental failure was flagged and rejected based onthe level of BVDV RNA in each well. The level of HCV RNA in each wellwas calculated according to a standard curve run in the same PCR plate.The percentage of inhibition or decrease of HCV RNA level due tocompound treatment was calculated using the DMSO or no compound controlas 0% of inhibition. The IC50 (concentration at which 50% inhibition ofHCV RNA level is observed) was calculated from the titration curve ofany given compound.

Example 5 HCV Ki Assay Protocol

HPLC Microbore Method for Separation of 5AB Substrate and Products

Substrate:

-   NH₂-Glu-Asp-Val-Val-(alpha)Abu-Cys-Ser-Met-Ser-Tyr-COOH

A stock solution of 20 mM 5 AB (or concentration of your choice) wasmade in DMSO w/0.2M DTT. This was stored in aliquots at −20 C.

Buffer: 50 mM HEPES, pH 7.8; 20% glycerol; 100 mM NaCl

Total assay volume was 100 μL

X1 conc. in (μL) assay Buffer 86.5 see above 5 mM KK4A 0.5 25 μM 1 M DTT0.5 5 mM DMSO or inhibitor 2.5 2.5% v/v 50 μM tNS3 0.05 25 nM 250 μM 5AB20 25 μM (initiate)

The buffer, KK4A, DTT, and tNS3 were combined; distributed 78 μL eachinto wells of 96 well plate. This was incubated at 30 C for ˜5-10 min.

2.5 μL of appropriate concentration of test compound was dissolved inDMSO (DMSO only for control) and added to each well. This was incubatedat room temperature for 15 min.

Initiated reaction by addition of 20 μL of 250 μM 5AB substrate (25 μMconcentration is equivalent or slightly lower than the Km for 5 AB).

-   -   Incubated for 20 min at 30 C.    -   Terminated reaction by addition of 25 μL of 10% TFA    -   Transferred 120 μL aliquots to HPLC vials

Separated SMSY product from substrate and KK4A by the following method:

Microbore Separation Method:

-   Instrumentation: Agilent 1100-   Degasser G1322A-   Binary pump G1312A-   Autosampler G1313A-   Column thermostated chamber G1316A-   Diode array detector G1315A-   Column:-   Phenomenex Jupiter; 5 micron C18; 300 angstroms; 150×2 mm; P/O    00F-4053-B0-   Column thermostat: 40 C-   Injection volume: 100 μL-   Solvent A=HPLC grade water+0.1% TFA-   Solvent B=HPLC grade acetonitrile+0.1% TFA

Time Flow Max (min) % B (ml/min) press. 0 5 0.2 400 12 60 0.2 400 13 1000.2 400 16 100 0.2 400 17 5 0.2 400

-   Stop time: 17 min-   Post-run time: 10 min.

Table 1 below depicts Mass Spec. (M−H, M+H, obs=observed), HPLC, ¹H-NMR(“Yes” if spectral data obtained), Ki, and IC₅₀ data for certaincompounds of the invention.

Compounds with Ki's ranging from 1 μM to 5 μM are designated A.Compounds with Ki's ranging from 1 μM to 0.5 μM are designated B.Compounds with Ki's below 0.5 μM are designated C. Compounds with IC₅₀'sranging from 1 μM to 5 μM are designated A. Compounds with IC₅₀'sranging from 1 μM to 0.5 μM are designated B. Compounds with IC₅₀'sbelow 0.5 μM are designated C.

TABLE 1 MS+ MS− HPLC, ¹H-NMR Compound (obs) (obs) R_(t)(min) Ki IC₅₀(CDCl₃) 1a 801.2 799.2 4.80 C C Yes 2a 744.0 742.2 4.0 C C Yes 3a 704.4702.6 3.63 C C Yes 4a 761.1 759.3 3.77 C C Yes 5a 744.2 742.1 4.05 C —Yes 6a 744.2 742.3 3.94 C — Yes 7a 730.2 728.3 3.90 C C Yes

All of the documents cited herein, are incorporated herein by reference.

We claim:
 1. A compound selected from the group comprising: